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 7. Nudt21 upregulates let-7-mediated HMGA2 silencing through APA. (A, 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. (A) The short and long HMGA2 3′-UTR transcripts were detected by RT-PCR. The long transcript-to-total transcript ratios are shown. (B) The short (Proximal) and long (Distal) HMGA2 3′-UTR transcripts were detected by 3′-RACE. (C) hTSCs were transfected with 100 ng psiCHECK2-HMGA2 along with shNudt21-1, shNudt21-2 or a scrambled control shRNA (control) and 0, 10 or 20 μg let-7 as indicated in the presence or absence of 0.5 mM H2O2. The luciferase activity was determined 48 h after transfection. (D) hTSCs were transfected with 100 ng psiCHECK2-HMGA2 or psiCHECK2-HMGA2 MUT along with 0, 10 or 20 μg let-7 as indicated in the presence or absence of 0.5 mM H2O2. The luciferase activity was determined 48 h after transfection. The data shown are from three replicates and are indicated as mean ± SD. *p < 0.05, **p < 0.01.