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. 2009 Mar 25;100(7):1275–1283. doi: 10.1111/j.1349-7006.2009.01166.x

Figure 2.

Figure 2

Figure 2

Induction of senescence and apoptosis by p21 overexpression. (a) Ad‐p21‐N‐ and Ad‐p21‐F‐infected HeLa cells were enlarged, flattened, and were senescence‐associated β‐galactosidase (SA‐β‐gal)‐positive. No change in morphology was observed in cells transfected with the mock vector and Ad‐p21C. Bar = 10 µM (b) The populations of SA‐β‐gal‐positive cells (in HeLa, DLD‐1, SKOV3, LoVo, and HCT116 cells) after 48 h of infection are shown. Data represent the average of three independent experiments and SD are indicated by error bars. (c) When LoVo and HCT116 cells were transfected with 20 MOI Ad‐p21N and Ad‐p21F, cells were enlarged, flattened, and had increased SA‐β‐gal positivity. At 50 MOI (LoVo) and 40 MOI (HCT116), cells were detached from the plate and floating. The small panel on the lower right for 40 or 50 MOI shows positive cells with the TUNEL assay. Mock‐transfected cells showed no change in morphology and were not positive for the TUNEL assays. The populations of apoptotic cells in LoVo and HCT116 infected with 50 or 40 MOI Ad‐p21 after infection of 48 h are shown in right graph as indicated by the significant increase of subG1 fraction. Data represent the average of three independent experiments and SD are indicated by error bars. Bar = 10 µM (d) Western blot of p21 expression levels in LoVo cell lysates from mock‐, Ad‐p21C‐, Ad‐p21N‐, and Ad‐p21F‐transfected cells with different MOIs. (e) Western blot of p53 expression levels in LoVo and HeLa cell lysates from mock‐, Ad‐p21C‐, Ad‐p21N‐ and Ad‐p21F‐transfected cells with different MOIs.