Human LIM1 Antibody

LIM1 in Human Brain (Cerebellum). LIM1 was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using Mouse Anti-Human LIM1 Monoclonal Antibody (Catalog # MAB2725) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in Purkinje neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human LIM1 by Western Blot LIM1 promoted CREB phosphorylation in EC. (A) Volcano plot of DEGs in LIM1-KD (#1 and #2) cells versus Ctrl cells generated from RNA-seq results. (B) The highly ranked GO cluster among commonly downregulated genes in LIM1-KD cells compared with Ctrl cells by Ingenuity Pathway Analysis (IPA). (C) Western blotting of LIM1, ERK, phospho-ERK, CREB, phospho-CREB, and GAPDH using Ctrl cells and LIM1-KD cells. (D) Relative intensities of Akt, phospho-Akt, ERK, phospho-ERK, CREB, phospho-CREB bands by western blot analysis. Signals intensities were normalized to the GAPDH intensity in the same samples and then the intensity of the LIM1-KD (#1 and #2) cells were normalized to Ctrl cells [n = 5 (ERK, p-ERK, CREB, p-ERK), n = 4 (Akt, p-Akt) one-way ANOVA]. (E) Immunofluorescent staining for phospho-CREB and human vimentin in sections of xenograft tumors derived from Ctrl cells and LIM1-KD cells. Red: phosphorylated CREB, blue: nucleus, green: human vimentin. (F) Intensity of phospho-CREB in the tumor region (n = 4, one-way ANOVA). *p < 0.05, **p < 0.01. Graphs show means ± standard deviations. (G) MTT assay to assess cell proliferation. HEC50B cells were treated with CREB inhibitors, KG501 and compoun3i, respectively. Red, orange, green and blue indicate inhibitor concentrations at 0.5 μM, 1.0 μM, 5.0 μM and 10 μM, respectively. (n = 5, two-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36969081), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human LIM1 by Western Blot LIM1 promoted CREB phosphorylation in EC. (A) Volcano plot of DEGs in LIM1-KD (#1 and #2) cells versus Ctrl cells generated from RNA-seq results. (B) The highly ranked GO cluster among commonly downregulated genes in LIM1-KD cells compared with Ctrl cells by Ingenuity Pathway Analysis (IPA). (C) Western blotting of LIM1, ERK, phospho-ERK, CREB, phospho-CREB, and GAPDH using Ctrl cells and LIM1-KD cells. (D) Relative intensities of Akt, phospho-Akt, ERK, phospho-ERK, CREB, phospho-CREB bands by western blot analysis. Signals intensities were normalized to the GAPDH intensity in the same samples and then the intensity of the LIM1-KD (#1 and #2) cells were normalized to Ctrl cells [n = 5 (ERK, p-ERK, CREB, p-ERK), n = 4 (Akt, p-Akt) one-way ANOVA]. (E) Immunofluorescent staining for phospho-CREB and human vimentin in sections of xenograft tumors derived from Ctrl cells and LIM1-KD cells. Red: phosphorylated CREB, blue: nucleus, green: human vimentin. (F) Intensity of phospho-CREB in the tumor region (n = 4, one-way ANOVA). *p < 0.05, **p < 0.01. Graphs show means ± standard deviations. (G) MTT assay to assess cell proliferation. HEC50B cells were treated with CREB inhibitors, KG501 and compoun3i, respectively. Red, orange, green and blue indicate inhibitor concentrations at 0.5 μM, 1.0 μM, 5.0 μM and 10 μM, respectively. (n = 5, two-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36969081), licensed under a CC-BY license. Not internally tested by R&D Systems.