Anti zeb1 antibody

We used ChIP coupled with quantitative PCR (qPCR) to characterize protein–DNA interaction. ChIP assays were performed as directed by us previously^{57 (link)} using a kit from Beyotime (Haimen, China). In brief, H9C2 cells were grown in a 10-cm dish. Chromatin was crosslinked with 1% formaldehyde. Cells (5 × 10⁶) were then lysed with 500 μl of ChIP lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl, pH 8.0) with a cocktail of protease inhibitors (Calbiochem) for 10 min on ice. The samples were sonicated to yield DNA fragments with an average size of ~200 bp. Protein–DNA complexes were immunoprecipitated with IgG or an anti-ZEB1 antibody (2 μg; Proteintech). The immunoprecipitated material was washed and heated at 65 °C overnight to reverse the crosslink. The ChIP DNA was column-purified (Qiagen, Valencia, CA, USA), and used for qPCR.

Protein extraction and western blotting analysis were performed using previously standard procedures (17 (link)). The following antibodies were used for western blotting: anti-E-cadherin antibody(#20874-1-AP Proteintech, China), anti-N-cadherin antibody(#22018-1-AP Proteintech, China), anti-ZEB1 antibody(#66279-1-Ig Proteintech, China), anti-Vimentin antibody(10366-1-AP Proteintech, China), anti-SOX2 antibody(#11064-1-AP Proteintech, China), anti-GAPDH antibody(#10494-1-AP Proteintech, China), and anti-PD-L1 antibody(#66248-1-Ig Proteintech, China).