anti human collagen type iv  (Thermo Fisher)

Journal: Life

Article Title: Lysophosphatidic Acid Modulates TGF-β2-Induced Biological Phenotype in Human Conjunctival Fibroblasts

doi: 10.3390/life14060770

Figure Lengend Snippet: LPA-induced effects on gene expression of ECM proteins COL1 , COL4 , COL6 , FN , and αSMA in 2D and 3D cultured HconF cells. The 2D and 3D HconF cells that had been incubated with 5 ng/mL TGF-β2 and/or 500 nM LPA in addition to non-treated control cells were subjected to qPCR analysis. Measurements were conducted in triplicate using fresh preparations (2D: n = 5; 3D: n = 10 spheroids each). * p < 0.05, ** p < 0.01, and *** p < 0.005 (ANOVA followed by Tukey’s multiple comparison test).

Article Snippet: Briefly, 4% paraformaldehyde-fixed 3D spheroids were incubated in 3% BSA in PBS for 3 h. After washing twice with PBS for 30 min each time, 3D spheroids were sequentially exposed to 1:200 dilution of primary antibodies, including an anti-human COL1 (#600-401-103-0.1, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL4 (#600-401-106S, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL6 (#009-001-108, ROCKLAND Antibodies & Assays, Limerick, PA, USA), and FN (#A-11, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) rabbit antibody at 4 °C overnight.

Techniques: Gene Expression, Cell Culture, Incubation, Control, Comparison

Journal: Life

Article Title: Lysophosphatidic Acid Modulates TGF-β2-Induced Biological Phenotype in Human Conjunctival Fibroblasts

doi: 10.3390/life14060770

Figure Lengend Snippet: Immunolabeling of ECM proteins COL1 , COL4 , COL6 , FN , and αSMA of the 3D HconF spheroids. The 3D HconF spheroids that had been incubated with 5 ng/mL TGF-β2 and/or 500 nM LPA in addition to non-treated control cells were subjected to immunocytochemistry analysis. Analyses were conducted in triplicate using fresh preparations ( n = 5). Relative staining intensities of the molecules are plotted in the right panels. * p < 0.05 (ANOVA followed by Tukey’s multiple comparison test).

Article Snippet: Briefly, 4% paraformaldehyde-fixed 3D spheroids were incubated in 3% BSA in PBS for 3 h. After washing twice with PBS for 30 min each time, 3D spheroids were sequentially exposed to 1:200 dilution of primary antibodies, including an anti-human COL1 (#600-401-103-0.1, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL4 (#600-401-106S, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL6 (#009-001-108, ROCKLAND Antibodies & Assays, Limerick, PA, USA), and FN (#A-11, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) rabbit antibody at 4 °C overnight.

Techniques: Immunolabeling, Incubation, Control, Immunocytochemistry, Staining, Comparison