Fig. 4

Overexpression of TZAP in early-passage MSCs decreased proliferation and differentiation potential. a The mRNA levels of TZAP and senescence-associated markers (p21 and p16) in pMSCs were assessed by qRT-PCR after transduction with control plasmids (CTR) or TZAP overexpression vectors (TZAP-OE). b Protein levels of TZAP, P16INK14 (P16) and P21 in pMSCs were compared through western blot analysis after TZAP overexpression. c The effect of overexpression of TZAP on the cell growth rate. d Bar = 100 μm. The EdU incorporation rate of pMSCs with TZAP overexpression compared to that of the controls. e Bar = 100 μm. Differentiation of pMSCs with TZAP overexpression was assessed by oil red O staining after adipogenic induction or by alizarin red S staining after osteogenic induction. f The relevant expression levels of bone markers (Alp, Runx2, and Col1a1) and adipogenic markers (Cebpa) were assessed by qRT-PCR and compared between pMSCs with TZAP overexpression and the control cell line. g PPARγ expression was examined by western blotting upon TZAP overexpression in pMSCs. h Bar = 100 μm. SA-β-gal staining of pMSCs with TZAP overexpression compared to that of the control. All data are represented as the mean ± SEM. n = 3. *p < 0.05; **p < 0.01; ^#p < 0.001, tested by one-way ANOVA