Anti srebp 1c sc 13551

We used the following commercially available antibodies: The anti-AdipoR1 (ab126611), anti-FN (ab2413), anti-Collagen I (ab34710), anti-AMPKα1 + AMPKα2 (ab131512), anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) (ab23875), anti-RAB7(ab50533) and anti-ADRP (ab52356) antibodies were from Abcam; the anti-phospho-AMPKα(Thr172) (2535), anti-mTOR(2972), anti-phospho-mTOR(Ser2448)(5536), anti-ULK1 (8054), anti-phospho-ULK1 (Ser555) (5869), anti-SQSTM1/p62(5114) and anti-LC3B (3868) antibodies were from Cell Signaling Technology; and the anti-SQSTM1/P62 (18420-1-AP), anti-LC3B (18725-1-AP), anti-Beclin1 (11306-1-AP), anti-ADRP (15294-1-AP), anti-ATG5 (10181-2-AP), anti-TFEB (13372-1-AP), anti-β-actin (60008-1-Ig), anti-GAPDH (60004-1-Ig) and anti-Lamin B(12987-1-AP) antibodies were from Proteintech. The anti-SREBP-1c(sc-13551) was from Santa Cruz Biotechnology. AdipoRon and CQ was obtained from Selleckchem. AICAR was provided by Apexbio Technology, SBI-0206965 was purchased from Sigma-Aldrich.

Chromatin was prepared from liver tissue or HepG2 cells. Chromatin was ultrasonically sheared using a Bioruptor UCD-200 (Diagenode, Belgium), and then immunoprecipitated with anti-H3k9Ac (9649 s, CST), anti-SREBP-1c (sc-13551, Santa Cruz). Normal rabbit IgG (B900610, Proteintech) was used as a mock antibody for negative control. After immunoprecipitating, reverse transcription quantitative polymerase chain reaction (qPCR) was performed. The primers were designed to amplify the reported SREBP-1c binding region (SRE) in mouse and human PNPLA3 promoter [10 (link), 11 (link)]. The primer sequences are as follow: mouse: forward 5’-CCTCCCACTGGCTTATTTGC-3’, reverse: 5’-CCGGTGTTGCTGCTCTGAG-3’, human: forward 5’- GCCATCGCCCTCCC AG-3’, reverse: 5’-GGGTGGGGACGACGT-3’. The recruitment is expressed as fold enrichment over IgG (mock).