Figure 4 | Cell Death & Disease

Figure 4

From: Notch activation enhances mesenchymal stem cell sheet osteogenic potential by inhibition of cellular senescence

Figure 4

Hes1 is required for JAG1-induced inhibition of MSC agin. (a) Bright field image showed a high infection efficiency in lentiviral GFP control MSCs after 24 h of culture. Scale bar represents 50 μm. (b) Western blot analysis confirmed the decrease of Hes1 protein levels at day 5 in Hes1 knockdown MSCs, even with JAG1 treatment. (c) SA-β-gal staining was performed to determine the extent of senescence in shHes1-infected MSCs at day 5 in cell sheet culture with JAG1 treatment. Scale bar represents 100 μm. (d) SA-β-gal positive cells in cell sheet cultures were quantitated by ImageJ. (e) Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) data showed that the decreased expression of p16 by JAG1 was rescued by knockdown of Hes1 in MSCs at day5 in sheet cultures. No changes were observed in p21 expression in 5-day sheet cultures. (f) Western blot data showed a similar expression pattern of p16 and p21 protein level to RNA expression in JAG1 treated wild type MSCs and Hes1 deficient MSCs. Results are mean±S.D. All experiments were performed in triplicate. *P<0.05 versus lgG control cells

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