FIGURE 4.

MYCL1 is a novel CXXC5 target. (A,B) LX-2 cells were treated with or without TGF-β1 (2 ng/ml). The cells were harvested at indicated time points and MYCL1 expression levels were examined by qPCR and Western. (C,D) C57/B6 mice were injected with or without CCl₄ for 4 weeks as described in section “Materials and Methods.” Primary hepatic stellate cells were isolated and CXXC5 expression levels were examined by qPCR and Western. N = 6 mice for each group. (E,F) C57/B6 mice were subjected to the BDL or the sham procedure for 2 weeks as described in section “Materials and Methods.” Primary hepatic stellate cells were isolated and CXXC5 expression levels were examined by qPCR and Western. N = 6 mice for each group. (G,H) LX-2 cells were infected with infected with adenovirus carrying CXXC5 expression vector (Ad-CXXC5) or GFP (Ad-GFP) followed by TGF-β1 (2 ng/ml) treatment for 48 h. MYCL1 expression was examined by qPCR and Western. (I) LX-2 cells were treated with or without TGF-β1 (2 ng/ml). The cells were harvested at indicated time points and ChIP assay was performed with anti-CXXC5 or IgG. (J) LX-2 cells were treated with or without TGF-β1 (2 ng/ml). The cells were harvested at indicated time points and ChIP assay was performed with anti-5′-methylcytosine, anti-acetyl H3K9, or anti-acetyl H3K27. (K) LX-2 cells were treated with or without TGF-β1 (2 ng/ml). The cells were harvested at indicated time points and ChIP assay was performed with anti-RNA Pol II (ser5) and anti-RNA Pol II (ser2). All experiments were repeated three times and one representative experiment is shown. *p < 0.05.