Suppression of pro-fibrotic responses in TGF-β1-stimulated LX2 cells via disruption of AKT interaction by p300 inhibitor. (A) Western blot analysis of p300 and αSMA in TGF-β1-stimulated LX2 cells with or without p300 inhibitors. (B) RT-qPCR analysis of expression of p300 and αSMA in TGF-β1-stimulated LX2 cells with or without p300 inhibitors. (C) Representative photomicrographs and percentages of Sirius red staining in TGF-β1-stimulated LX2 cells with or without p300 inhibitors (n = 5). (D) Western blot analysis of fibrosis marker proteins, COL1A, CTGF, FN, TNC, and Periostin, in TGF-β1-stimulated LX2 cells with or without p300 inhibitors. (E) RT-qPCR expression analysis of fibrosis marker genes, COL1A1, COL3A1, CTGF, and FN in TGF-β1-stimulated LX2 cells with or without p300 inhibitors. (F) ChIP assay using IgG or anti-p300 antibody in TGF-β1-stimulated LX2 cells with or without p300 inhibitors. Data are represented as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by ordinary one-way ANOVA test. (G) Immunoprecipitation assay using anti-HA or anti-Myc antibodies in LX2 cells (co-transfected Myc-AKT and HA-AKT). β-actin was used as the sample loading control.
