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 1. HMGA2 overexpression protects hTSCs from H2O2-induced loss of self-renewal capacity and pluripotency. hTSCs transfected with HMGA2 overexpression lentivirus (HMGA2 OE) or the negative control (nc) were subjected to H2O2 treatment at indicated concentrations for 24 h. Non-transfected cells (mock) were included for comparison. (A, B) The HMGA2 mRNA (a) and protein (b) levels were determined by qRT-PCR and western blot analysis, respectively. (C) Clonogenicity was evaluated by the colony formation assay. (D) The quantitative expression of stemness markers Nanog, Nucleostemin, Oct-4, and SSEA-4 of treated hTSCs were measured by qRT-PCR. (E) The multilineage differentiation capability was assessed as described in materials and methods. Scale bar = 50 μm. (F) The expression of markers for lineage-specific differentiation (PPARγ for adipogenesis, Runx-2 for osteogenesis and Sox-9 for chondrogenesis) at day 0 and day 14 of differentiation was evaluated by qRT-PCR. The data shown are from three replicates and are presented as mean ± SD. *p < 0.05, **p < 0.01.