Research Paper Volume 8, Issue 8 pp 1670—1689

Figure 2. BMI1 regulates cellular senescence in hUCB-MSCs. (A-C) To assess the role of BMI1 in hypoxia, hypoxic-cultured hUCB-MSCs were transfected with Luc control and shBMI1, and the senescence phenotype was investigated. (A) Expression levels of BMI1 and p16^INK4a proteins were confirmed via western blot analysis. (B) To assess the proliferation of BMI1-down-regulated hUCB-MSCs, an MTT assay was performed. (C) The senescent state of cells was confirmed via SA-β-gal staining. (D-G) To confirm the effects of BMI1 on cellular senescence, normoxic-cultured hUCB-MSCs were induced to overexpress GFP and BMI1. (D) Western blot analysis was performed to confirm the expression levels of BMI1 and p16INK4a proteins. (E) MTT assay was conducted to evaluate the proliferation of BMI1-up-regulated hUCB-MSCs. (F) After the overexpression of BMI1, CPDLs were determined to evaluate the proliferative ability of the hUCB-MSCs. (G) After several passages, the senescent state of cells was confirmed via SA-β-gal staining. The results show 1 representative of 3 independent experiments. Error bars represent mean±s.e.m. from three separate experiments. ** P<0.01, *** P<0.005 using Student’s t-test.