Figure 3.

High autophagic flux preserves ESC identity. (a) Western blotting for ATG3 and LC3 in Atg3^+/+, Atg3^+/−, and Atg3^−/− ESCs. β-Actin served as the loading control. (b) Restore of wild-type but not V8D mutant Atg3 expression in Atg3^+/−, and Atg3^−/− ESCs compensated for reprogramming efficiency. (c) Quantification of AP-positive colony numbers in (b). Data shown are mean±S.D., n=3, *P<0.05, **P<0.01, ***P<0.001 (Student’s t-test). (d) Long-lived protein degradation in Atg3^+/+, Atg3^+/−, and Atg3^−/− ESCs under EBSS starvation conditions (5 h) with or without 3-MA treatment (10 mM, 5 h). Data shown are mean±S.D., n=3, **P<0.01, ***P<0.001 (Student’s t-test). (e) qPCR for pluripotency marker genes in ATG3 rescue ESC lines. Data shown are mean±S.D., n=3, *P<0.05, **P<0.01 (Student’s t-test). (f) Statistic analysis of the weight of teratomas formed by Atg3^+/+, Atg3^+/−, and Atg3^−/− ESCs. *P<0.05, ***P<0.001 (Student’s t-test). (g) Statistic analysis of the chimeric formation ratio of embryos (11.5 d) formed by Atg3^+/+, Atg3^+/−, and Atg3^−/− ESCs