For Western blot analysis of PAX6, 10 ug of whole cell lysates was loaded to 4-20% gradient SDS-PAGE gels and then transferred to nitrocellulose membranes (BIO-RAD) by using Trans-Blot® Turbo™ Transfer System (BIO-RAD). Blocked membranes were incubated with antibodies against PAX6 (Cat. no. 5790, Abcam) or GAPDH (Cat. no. 2118, Cell Signaling), respectively. For Western blot analysis of LDHA, undifferentiated hESC/hiPSC cells were sorted for hTRA- 1-81+ in order to control for the homogeneity of the cells, and then the rest of the procedure ensued as above. LDHA (Cat. no. 2012S, Cell Signaling), β-ACTIN (Cat. no. MA5-15739-HRP, Thermo Scientific).
