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. 1999 Oct 1;13(19):2514–2526. doi: 10.1101/gad.13.19.2514

Figure 5.

Figure 5

Effects of RIP cleavage on apoptosis. (A) HeLa cells were cotransfected with pRSV–LacZ and different RIP constructs with (open bars) or without CrmA (solid bars) as indicated. Twenty-four hours post-transfection, the cells were fixed and stained with X-gal. Viable blue cells, determined by their sizes and shapes, were counted from 20 randomly selected fields. Data shown represent three independent experiments. The difference of viable cells of wild-type RIP vs. RIP (D324K) transfection is statistically significant (P < 0.005). (B) Duplicates of MCF7 cells were cotransfected with pRSV–LacZ and different RIP constructs. Twenty-four hours post-transfection, half of the cells were treated with 15 ng/ml TNF for 18 hr and then fixed and stained with X-gal. Alive blue cells were counted from 20 randomly selected fields for each sample. Percentage of viable cells for each plasmid was calculated by dividing the number of viable transfected cells after treatment with the number of viable transfected cells before treatment. Data shown represent three independent experiments. The differences of viable cells of wild-type versus RIP (D324K) and RIPc versus vector transfections are statistically significant (both P < 0.05). (C) MCF7 cells were cotransfected with pRSV–LacZ and different RIP constructs as indicated. Cells were treated and counted as described in B. The difference of viable cells of wild-type vs. RIP (D324K) transfection is statistically significant (P < 0.005).