Research Paper Volume 12, Issue 18 pp 18436—18452

Figure 4. Autophagy blockage abolishes the protective effects of HMGA2 overexpression against H₂O₂-induced loss of potential regenerative power and pluripotency. hTSCs transfected with lenti-HMGA2 (HMGA2 OE) or the empty virus (nc) were incubated with 3-MA and rapamycin for 24 h, alone or in combination as indicated, and subsequently exposed to 0.5 mM H₂O₂ for 24 h. Untransfected cells (mock) were included for comparison. (A) Clonogenicity was assessed by the colony formation assay. (B) The quantitative expression of stemness markers Nanog, Nucleostemin, Oct-4, and SSEA-4 were measured by qRT-PCR. (C–E) The multilineage differentiation capability and the expression of markers for lineage-specific differentiation (PPARγ for adipogenesis, Runx-2 for osteogenesis and Sox-9 for chondrogenesis) was evaluated by qRT-PCR. Scale bar = 50 μm. The data shown are from three replicates and are indicated as mean ± SD. *p < 0.05, **p < 0.01.