Anti c ebpa

The plasmid encoding pAAV‐HBV1.2 was kindly provided by Pei‐Jer Chen from the Department of Internal Medicine, National Taiwan University Hospital, National Taiwan University College of Medicine. HBV promoters including X promoter (XP), core promoter (CP), PreS1 promoter (preS1P), PreS2 promoter (preS2P) luciferase report vectors (pGL3‐Xp, pGL3‐S1p, pGL3‐S2p and pGL3‐Cp) were created in our laboratory according to previous studies.³⁷ pHBV1.37 plasmid was generated in our laboratory according to previous studies.³⁷ The antibodies used were listed: anti‐p‐ERK (4370S, Cell Signaling Technology, USA), anti‐ERK (4695S, Cell Signaling Technology, USA), anti‐HNF1α (89670S,Cell Signaling Technology,USA), anti‐HNF4a(3113S, Cell Signaling Technology,USA), anti‐C/EBPa (8178S, Cell Signaling Technology, USA), anti‐PPARa (ab3484,abcam,USA), anti‐NEDD8 (ab81264, abcam, USA), anti‐GAPDH (2118S, Cell Signaling Technology, USA), anti‐Actin (A1015,DAWEN BIOTECH,CHINA) Normal Rabbit IgG (WD‐GAR007, DAWEN BIOTECH,CHINA),Normal Mouse IgG (GAM007,MULTI SCIENCES,CHINA).

Proteins were separated by SDS‐PAGE and transferred onto nitrocellulose membranes. The membranes were incubated overnight with primary antibodies. The antibodies we used include rabbit antibodies by Abcam (Cambridge, MA, USA): anti‐osterix (polyclonal), anti‐C/EBPa (monoclonal), anti‐β‐catenin (monoclonal), anti‐opsteopontin (monoclonal) and anti‐ALP (monoclonal); rabbit antibodies by Cell Signalling Technology (Danvers, MA, USA): anti‐PPARγ (monoclonal), anti‐LRP6 (monoclonal) and anti‐Phospho‐LRP6 (polyclonal); mouse mAb by MBL (Nagoya, Japan): anti‐Runx2; rabbit polyclonal antibodies by Proteintech (Wuhan, China): anti‐aP2, and anti‐β‐actin; rabbit polyclonal antibody by SAB: anti‐H3K9me2 and rabbit polyclonal antibody by Bioworld: anti‐H3K27me2. The membranes were then incubated with the corresponding horseradish peroxide‐labeled IgG (1:3000) for 2 hours. Chemiluminescence reagent (Advansta, Menlo Park, CA, USA) was finally used to visualize the results.