Total protein of cells was extracted with a radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor under the condition of an ice bath. The extracted protein and loading buffer were added and transferred onto 0.22-mm methanol-excited polyvinylidene fluoride membranes after SDS-PAGE electrophoresis, and then membranes were blocked with 5% nonfat milk. Next, we incubated membranes with the following primary antibodies at 4°C overnight: TAF15 (1:1,000) (Cell Signaling Technology, USA), MTF1 (1:1,000) (Proteintech, USA), YY2 (1:200) (Santa Cruz Biotechnology, USA), GTSE1 (1:1,000) (Proteintech, USA), and GAPDH (1:10,000) (Proteintech, USA), followed by incubation with related horseradish peroxidase (HRP)-conjugated secondary antibodies, including goat anti-rabbit or goat anti-mouse secondary antibody (1:10,000) (Proteintech, USA) at room temperature for 1.5 h. The relative integrated density values (IDVs) were observed based on GAPDH as a control.
