Fig. 5. ATF3 translocates to the nucleus of LX-2 cells and promotes the transcription of pro-fibrogenic genes.

a LX-2 cells were treated with 10 ng/ml TGF-β for 48 h, and the expression and location of ATF3 and COL1 were determined by confocal microscopy. DAPI-stained nuclei blue; scale bar, 20 μm. b, c The RNA and protein were extracted from the nuclei or cytoplasm of LX-2 cells treated with or without TGF-β, the expression of ATF3 was assessed by qRT-PCR and western blot. GAPDH was used as control in cytoplasm and H3K27 as nuclear. d LX-2 cells were treated with TGF-β and ChIP analyses were performed on indicated genes promoter regions, using anti-ATF3 antibody. Enrichment was shown relative to input. e ATF3 and SMAD3 antibodies were used for co-immunoprecipitation (IP) with LX-2 lysates treated with or without TGF-β. The data were expressed as the mean ± SEM for at least triplicate experiments. GAPDH was used as an internal control; */#p < 0.05.