Research Paper Volume 12, Issue 18 pp 18436—18452

Nudt21-mediated alternative polyadenylation of HMGA2 3′-UTR impairs stemness of human tendon stem cell

Figure 3. HMGA2 overexpression activates autophagy in H2O2-treated hTSCs. (A) hTSCs were incubated with rapamycin and 3-MA for 24 h, alone or in combination as indicated, and subsequently exposed to 0.5 mM H2O2 for 24 h. hTSCs were transfected with lenti-HMGA2 (HMGA2 OE) or the negative control (nc) in the presence or absence of 3-MA for 24 h, and subsequently exposed to 0.5 mM H2O2 for 24 h. Normal cells (mock) were included for comparison. The protein levels of the autophagic markers LC3-I, LC3-II, and Beclin-1 were determined by western blot analysis. (B) GFP-LC3-transfected hTSCs were subjected to treatments as described in a. GFP-LC3-labeled vacuoles (puncta) were detected by fluorescence imaging at 60× magnification. The data shown are from three replicates and are presented as mean ± SD. *p < 0.05, **p < 0.01.