Sc 17759

Staining reagents included CtBP2 (sc-17759; 1:200; Santa Cruz Biotechnology, Inc.) and TGIF (ab52955; 1:1,000; Abcam) primary antibodies, anti-mouse (SAB3701092) or anti-rabbit (F4890) secondary antibodies (MilliporeSigma), and Hoechst 33342 (Beyotime Institute of Biotechnology). The cells were digested with trypsin and pipetted vigorously to make a single-cell suspension. The cells were placed on glass coverslips (24-well plates), incubated at 37°C and fixed with 4% PFA for 20 min at 15–25°C. The glass coverslips were then washed three times with PBS for 10 min each and blocked with 10% bovine serum albumin (BSA) for 2 h at 15–25°C. Subsequently, all cells were incubated with primary antibody (1:200) overnight at 4°C. The following day, the cells were incubated with secondary antibody (1:2,000) for 2 h at 15–25°C. Finally, the cells were stained with Hoechst 33258 (Beyotime Institute of Biotechnology) at room temperature for 10 min, fixed with anti-fade solution and imaged under a fluorescence microscope (Zeiss AG).