Human Laminin  alpha 4 Antibody

Detection of Human Laminin alpha 4 by Simple Western^TM.

Simple Western lane view shows lysates of human placenta tissue, loaded at 0.2 mg/mL. A specific band was detected for Laminin a4 at approximately 233 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Adhesion Induced by Laminin alpha 4 and Neutralization by Human Laminin alpha 4 Antibody.

Recombinant Human Laminin a4 (Catalog # 7340-A4) supports adhesion in the HT1080 human fibrosarcoma cell line in a dose-dependent manner (orange line), as measured by Calcein AM. Adhesion elicited by Recombinant Human Laminin a4 (5 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340). The ND₅₀ is typically 1.5-7.5 µg/mL.

Detection of Human Laminin alpha 4 by Western Blot

Flow induces remodeling of the endothelial extracellular matrix.A, Proteome and (B) cell surface proteome data of identified laminins. T or CT represents the enzyme used for protein digestion. The depicted numbers represent the biotin-modified lysine residue. C, Schematic representation of laminin 411 with the linker region between domains LG3 and LG4. The blue lysine residues were found to be decreased under flow conditions (as depicted in panel B). D, Immunoblot analysis of LAMA4 in ECs exposed to flow over time. alpha -tubulin was used as a loading control. E, Protein distribution of LAMA5 and LAMA4 after flow-exposure. Green represents LAMA4 or LAMA5, blue is HOECHST. The scale bar is 10 μm. Depicted are maximum intensity projections. F, Proteome and (G) cell surface proteome data of identified fibulins. H, Immunofluorescence analysis of EFEMP1 and FBLN2 abundance and distribution affected by flow. Red is VE-cadherin, blue is HOECHST. Depicted are maximum intensity projections. Micrographs are representative for 3 independent experiments. The scale bar is 10 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32332107), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Immunohistochemistry

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Immunohistochemistry

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

Knockdown of LAMA4 suppresses the malignant behaviour of osteosarcoma and accelerates ER stress-induced apoptosis. A The expression levels of LAMA4 in osteosarcoma (n = 88) and normal tissues (n = 564) were analysed from the TNMplot database. B LAMA4 was knocked down in MNNG/HOS and 143B cells. The expression of LAMA4 was detected by Western blot. GAPDH was used as the loading control. C-D The cells (3000 cells/well) with or without LAMA4 depletion were tested for cell growth in the colony formation assay. Viable colonies after 1 week were counted and are shown (C). Data are depicted as bar graphs (D). E-F The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (E). Data are depicted as bar graphs (F). G-H MNNG/HOS cells with or without LAMA4 knockdown were treated with 3 μM TM for 36 h. Cell apoptosis and cell viability were analysed by Western blot (G) and CCK8 assays (H). GAPDH was used as the loading control. I-J MNNG/HOS cells with or without LAMA4 knockdown were treated with 1 μM TG for 36 h. Cell apoptosis and cell viability were analysed by Western blot (I) and CCK8 assays (J). GAPDH was used as the loading control. K-M MNNG/HOS cells (106 cells per mouse) with or without LAMA4 knockdown were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumours (K). The weights of the tumours were calculated and analysed (L). The expression levels of PARP, cleaved PARP and LAMA4 in tumours were measured by Western blot (M). Data in D, F, H, J, L were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Laminin alpha 4 by Western Blot

KLF4 upregulates LAMA4 expression. A-B Heatmap showing the altered genes identified by RNA sequencing analysis in LINC00629 (A)- or KLF4 (B)-depleted MNNG/HOS cells. C Volcano plots showing differentially expressed mRNAs for LINC00629 knockdown or KLF4 knockdown versus the controls (absolute log2-fold change > 1 and q value < 0.05). The green dot indicates the downregulated genes, and the red dot indicates the upregulated genes. D The mRNA levels of LAMA4 are shown from RNA sequencing data. E-F The protein and mRNA levels of LAMA4 were detected by Western blot (E) and qRT–PCR (F) in MNNG/HOS cells with or without LINC00629 or KLF4 knockdown. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. G MNNG/HOS cells with or without KLF4 or LINC00629 knockdown were treated with 3 μM TM for 36 h. The expression of LAMA4 was detected by Western blot. Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. H-I LINC00629 was overexpressed in MNNG/HOS cells with or without KLF4 knockdown. The protein and mRNA levels of LAMA4 were detected by Western blotting (H) and qRT–PCR (I). Numbers represent the relative intensities of western blot bands of LAMA4 to GAPDH. Data in F and M were analysed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36539799), licensed under a CC-BY license. Not internally tested by R&D Systems.