Srebp2 antibody

Deoxycholic acid and JTE-013 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Triamterene, 4-DAMP, ketoconazole, lovastatin, KG-501 and fatostatin were purchased from MedChemexpress (Monmouth Junction, NJ, USA). Methoctramine was from AdooQ Bioscience (Nanjing, China). Z-Guggulsterone was from Santa cruz Biotechnology (Santa cruz, CA, USA). SREBP2 antibody was obtained from R&D Systems (Minneapolis, MN, USA).

ChIP assay was performed with a kit (Beyotime, Shanghai, China) according to the manufacturer’s protocols. EL4 cells were treated as indicated and cross-linked by 1% formaldehyde. Nuclei were collected and subjected to sonication. Then the chromatin extracts were pre-cleared with protein A + G agarose and immunoprecipitated with SREBP2 antibody (#AF7119, R&D Systems, MN, USA) or rabbit IgG (#AC005, ABclonal, Wuhan, China) overnight (4 °C). De-crosslinking was implemented followed by DNA purification. Finally, PCR amplification was performed using primers (forward: 5′-GAA GGG CTG GTC TCA CAA AG-3′, and reverse: 5′-CGA AGG CGC TCT GTG ATT G-3′) encompassing the predicted SREBP2 binding region of murine Cyp51 promoter.

Colon cancer cells were harvested and detergent-solubilized cell lysates were obtained as described previously^{41 (link)–44 (link)}. Equal amounts of cell lysates were resolved by SDS-PAGE and subjected to western blot analysis. The SREBP1 antibody was from BD Biosciences (CA, USA), whereas the SREBP2 antibody was from R&D Systems (MN, USA). Note that the SREBP antibodies recognize both the precursor and mature active forms of the respective proteins. The γ-tubulin antibody was from Sigma-Aldrich.