Figure 3.

Inhibition of B‐myb expression promoted HAECs undergoing premature senescence and upregulating p53 and p21 expression. A, HAECs were transfected with siRNA against B‐myb (siB‐myb) or the controls (siNC) for 3 d. The knockdown efficiencies of B‐myb mRNA and protein levels were confirmed by real‐time PCR and western blotting. GAPDH was used for normalization. The mRNA data are presented as mean ± SEM of three independent experiments. ** indicates P<.01 compared with the control. A typical group of blots was shown from one of three independent experiments. GAPDH was used as a loading control. B, The percentage rate of SA‐β‐gal‐positive cells and the expression protein levels of p53, p21, p16 and pRb were analysed after the cells were transfected with siB‐myb or siNC for 7 d. The SA‐β‐gal‐positive cells were observed under inverted microscope (×100 magnification) after the treated cells were stained with senescence‐associated β‐galactosidase (SA‐β‐gal). A representative group image of stained cultures is shown. The percentage rate of SA‐β‐gal‐positive cells was analysed. Results are presented as the mean ± SEM of three independent experiments. *** indicates P<.001 compared with the control. A typical group of blots for analysing the expression protein levels of p53 and p21 in the cells is shown from one of three independent experiments. GAPDH was used as a loading control