Human Lysyl Oxidase Homolog 2/LOXL2 Antibody

Detection of Human Lysyl Oxidase Homolog 2/LOXL2 by Western Blot

Pro-fibrotic signalling pathways in human lung fibroblasts.(A–C) Healthy lung fibroblasts exposed to control, EGF, TGF beta 1, DMOG, Wnt3 alpha or Wnt5 alpha signalling for 24, 48, or 72 hr. n = 3 independent experiments. (A) Protein expression of phospho-ERK, phospho-SMAD2/3, HIF1 alpha, and active beta -catenin at 24 hr of exposure to conditions. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (B) LOXL2 and PLOD2 protein levels at 24, 48, or 72 hr of exposure to conditions. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (C) Expression of COL3A1 in healthy lung fibroblasts exposed to conditions for 24, 48, or 72 hr using the delta delta Ct method. Bars indicate geometric means. ****p < 0.0001 by Dunnett’s multiple comparisons test. (D) Protein expression of HIF1 alpha, LOXL2, and PLOD2 in IPF fibroblasts exposed to control media or IOX2 for 24, 48, or 72 hr. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (E) Fold change in mRNA levels of LOXL2, PLOD2 and the HIF pathway activation marker gene carbonic anhydrase IX/9 (CA9) in MRC5 fibroblasts after incubation in nomoxia (21% O2) or hypoxia (1% O2) for 24 hr. beta -actin-normalised mRNA levels under nomoxia were used to set the baseline value at unity. Data are mean ± s.d. n = 3 samples per group. ****p < 0.0001 using unpaired t test. Figure 2—figure supplement 1—source data 1.Full membrane scans for western blot images for Figure 2—figure supplement 1a, b, d.Full membrane scans for western blot images for Figure 2—figure supplement 1a, b, d. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35188460), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Lysyl Oxidase Homolog 2/LOXL2 by Western Blot

HIF pathway activation regulates PLOD2 and LOXL2 expression in lung fibroblasts from patients with IPF.(A) Fold changes in mRNA levels of HIF1 alpha (HIF1A), HIF2 alpha (EPAS1), and HIF1 beta (ARNT) in primary human lung fibroblasts from patients with IPF transfected with indicated siRNA followed by treatment with DMOG. beta -actin-normalised mRNA levels in control cells were used to set the baseline value at unity. Data are mean ± s.d. n = 3 samples per group. (B, C) Fold change in mRNA levels of LOXL2 (B) and PLOD2 (C) in IPF fibroblasts transfected with indicated siRNA followed by treatment with DMOG or vehicle control. beta -actin-normalised mRNA levels in control cells were used to set the baseline value at unity. Data are mean ± s.d. n = 3 samples per group. ns (not significant, p > 0.05); *p < 0.05; ****p < 0.0001 by Dunnett’s multiple comparisons test. (D) PLOD2, LOXL2 and HIF1 alpha and beta -tubulin protein levels in IPF fibroblasts transfected with indicated siRNA followed by treatment of DMSO or DMOG. beta -tubulin was used as a loading control. The full blots are shown in Figure 3—source data 1.Figure 3—source data 1.Full membrane scans for western blot images for Figure 3d.Full membrane scans for western blot images for Figure 3d. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35188460), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Lysyl Oxidase Homolog 2/LOXL2 by Western Blot

Pro-fibrotic signalling pathways in human lung fibroblasts.(A–C) Healthy lung fibroblasts exposed to control, EGF, TGF beta 1, DMOG, Wnt3 alpha or Wnt5 alpha signalling for 24, 48, or 72 hr. n = 3 independent experiments. (A) Protein expression of phospho-ERK, phospho-SMAD2/3, HIF1 alpha, and active beta -catenin at 24 hr of exposure to conditions. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (B) LOXL2 and PLOD2 protein levels at 24, 48, or 72 hr of exposure to conditions. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (C) Expression of COL3A1 in healthy lung fibroblasts exposed to conditions for 24, 48, or 72 hr using the delta delta Ct method. Bars indicate geometric means. ****p < 0.0001 by Dunnett’s multiple comparisons test. (D) Protein expression of HIF1 alpha, LOXL2, and PLOD2 in IPF fibroblasts exposed to control media or IOX2 for 24, 48, or 72 hr. beta -actin was used as a loading control. The full blots are shown in Figure 2—figure supplement 1—source data 1. (E) Fold change in mRNA levels of LOXL2, PLOD2 and the HIF pathway activation marker gene carbonic anhydrase IX/9 (CA9) in MRC5 fibroblasts after incubation in nomoxia (21% O2) or hypoxia (1% O2) for 24 hr. beta -actin-normalised mRNA levels under nomoxia were used to set the baseline value at unity. Data are mean ± s.d. n = 3 samples per group. ****p < 0.0001 using unpaired t test. Figure 2—figure supplement 1—source data 1.Full membrane scans for western blot images for Figure 2—figure supplement 1a, b, d.Full membrane scans for western blot images for Figure 2—figure supplement 1a, b, d. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35188460), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Lysyl Oxidase Homolog 2/LOXL2 by Western Blot

IOX2-mediated HIF pathway activation promotes PLOD2 and LOXL2 expression in the 3D in vitro model of fibrosis. Lung fibroblasts from IPF patients were used in the 3D model of fibrosis in the presence of IOX2 or vehicle control as indicated. Protein expression of (A) HIF1 alpha, PLOD2, and (B) LOXL2 following 2 weeks of culture in the presence or absence of TGF beta 1 with or without IOX2 (50 μM or 250 μM) or vehicle control. beta -actin loading control. Blots representative of experiments from two separate IPF donors. The full blots are shown in Figure 5—figure supplement 1—source data 1.Figure 5—figure supplement 1—source data 1.Full membrane scans for western blot images for Figure 5—figure supplement 1a, b.Full membrane scans for western blot images for Figure 5—figure supplement 1a, b. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35188460), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Lysyl Oxidase Homolog 2/LOXL2 by Western Blot

IOX2-mediated HIF pathway activation promotes PLOD2 and LOXL2 expression in the 3D in vitro model of fibrosis. Lung fibroblasts from IPF patients were used in the 3D model of fibrosis in the presence of IOX2 or vehicle control as indicated. Protein expression of (A) HIF1 alpha, PLOD2, and (B) LOXL2 following 2 weeks of culture in the presence or absence of TGF beta 1 with or without IOX2 (50 μM or 250 μM) or vehicle control. beta -actin loading control. Blots representative of experiments from two separate IPF donors. The full blots are shown in Figure 5—figure supplement 1—source data 1.Figure 5—figure supplement 1—source data 1.Full membrane scans for western blot images for Figure 5—figure supplement 1a, b.Full membrane scans for western blot images for Figure 5—figure supplement 1a, b. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35188460), licensed under a CC-BY license. Not internally tested by R&D Systems.