Anti e2f7

The chromatin immunoprecipitation assay was performed using a Pierce Magnetic ChIP Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. The antibodies used for immunoprecipitation were anti-H3K27ac (CST), anti-E2F7 (Abcam), and anti-RUNX1 (Abcam). Purified DNA was detected with specific primers for the promoter region using quantitative PCR. The primer sequences used for quantitative PCR are shown in Table S6.

RIPA lysis buffer including protease inhibitor (Madison, WI, USA) was utilized to isolate total protein. The proteins were separated by 10% SDS-PAGE and transferred onto a PVDF membrane (Millipore, NY, USA). Subsequently, the membrane was blocked with 5% skim milk for 1 h. Then the membrane was incubated with primary antibodies overnight at 4 °C, and hatched with secondary antibodies for 1 h at room temperature. With an ECL Kit (Millipore, NY, USA), the proteins were visualized and tested utilizing an ImageJ software (Bio-Rad, Hercules, CA). Western blotting analysis was executed utilizing the primary antibodies: anti-E2F7 (1: 1000, ab245655, Abcam, MA, USA), E-cadherin (1:1000, #14472, Cell Signaling Technology, Inc.), N-cadherin (1:1000, #13116, CST), Vimentin (1:1000, #5741, CST), Snail (1:1000, #3879, CST), GAPDH (1:5000, #5174, CST) and the secondary antibodies (1:5000, #7076 or #7074, CST). The expression of GAPDH was a control.