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 6. Nudt21 is induced by H2O2 and down-regulates HMGA2 in hTSCs. (A) hTSCs were subjected to H2O2 treatment at indicated concentrations for 24 h. The protein levels of Nudt21 were determined by western blot analysis. n = 3; *p < 0.05, **p < 0.01 vs. 0 mM H2O2. (B) hTSCs transfected with shNudt21-1, shNudt21-2 or a scrambled control shRNA (nc) lentivirus were cultured in the presence or absence of 0.5 mM H2O2 for 24 h. Nudt21 and HMGA2 protein levels were determined by western blot analysis. Untransfected cells (mock) were included for comparison. (CE) hTSCs transfected with a scrambled control shRNA (nc), shNudt21-2 and shHMGA2, alone or in combination as indicated were cultured in the presence or absence of 0.5 mM H2O2 for 24 h. (C) Clonogenicity was evaluated by the colony formation assay. (D) Cell viability was determined by the CCK-8 assay. (E) Cell senescence was assessed by β-gal staining 72 h after H2O2 treatment. The data shown are from three replicates and are indicated as mean ± SD. *p < 0.05, **p < 0.01.