Anti c jun sc1694

Cells were harvested and extracted with lysis buffer (Hepes 20 mM, pH 8; NaCl 150 mM; EDTA 2 mM; Nonidet P40 1%; SDS 0.1%; sodium deoxycholate 0.5% containing protease inhibitors (Complete mini EDTA free, Roche) and phosphatase (PhosSTOP, Roche). Lysates were centrifuged at 13,000 rpm for 15 min at 4°C, and supernatants collected. Equal amounts of protein were separated by SDS-PAGE, transferred to nitrocellulose membrane (Protran, Whatman) at 30 V overnight at 4°C and blocked with 4% skimmed milk for 2 h. The antibodies used in immunoblotting were as follows: Anti-pATF2 (pThr71) (sc-7982-R, Santa Cruz Biotechnology, Santa Cruz, CA); anti-ATF2 (sc-6233, Santa Cruz Biotechnology, Santa Cruz, CA); anti-c-Jun (sc1694, Santa Cruz Biotechnology, Santa Cruz, CA); anti-MMP9 (AV33090; Sigma); anti-ADAM19 (ARP49780_P050. Aviva Systems Biology); anti-Actin (I 19-sc1616, Santa Cruz Biotechnology, Santa Cruz, CA).

The cell culture reagents were obtained from Hyclone (Logan, USA) and the fetal bovine serum (FBS) was obtained from Gibco (New York, USA). The taurine, cholesterol, cholesterol esterase, cholesterol oxidase, horseradish peroxidase and sodium taurocholate hydrate were purchased from Sigma (St. Louis, MO, USA). The PowerOpti-ECL Western blotting detection reagent was purchased from GenView (Florida, USA). The primary (anti-CYP7A1, sc-25536; anti-c-jun, sc-1694; anti-p-c-jun, sc-7980-R; and anti–HNF4α, sc-8987) and secondary (anti-rabbit immunoglobulin G, sc-2004) antibodies were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary anti-MEK1/2 antibody (D1A5) was obtained from Cell Signaling Technology (Beverly, MA, USA). The primary anti-β-actin antibody was obtained from GenView (Florida, USA). The BCA Protein Assay Kit, Cell Lysis Buffer and PMSF were purchased from Beyotime (Nanjing, Jiangsu, China). All the other chemicals were purchased from Dingguo Changsheng Biotechnology Co. Ltd (Beijing, China).