FIGURE 5.
Expression of p14ARF in p53-deficient SAOS-2 cells triggers Mcl-1 and Bcl-xL deprivation and apoptosis. A, p53-deficient SAOS-2 osteosarcoma cells were infected with indicated adenoviral vectors at 50 m.o.i. or mock-treated (control). Total cellular proteins were separated by SDS-PAGE and subjected to Western blot analysis after 24, 48, and 72 h. B, SAOS-2 cells were treated as described in A. An N-terminal conformational change of Bak (BakNT*) was detected by flow cytometry using a conformation-specific antibody at 48 h after infection. C, cells were treated as described in A. Apoptotic cells were analyzed by flow cytometry to detect fragmented nuclear DNA 72 h after infection. D, expression of Bax was down-regulated by RNA interference (right panel, confirmation by Western blot analysis). 24 h later, cells were infected with the indicated adenoviral vectors (50 m.o.i.) or mock-treated for 48 h. Apoptotic DNA fragmentation was determined by flow cytometry. Error bars represent the means ± S.D. from triplicates.