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, Legends Figure 1: E2F1 induces apoptosis in lung adenocarcinoma cells through a caspase-8-and DNA-binding domain-dependent mechanism
, 1/GFP (open bars) or a pcDNA3.1/E2F1 (closed bars) vector. Apoptosis was monitored after 6 days of geniticin selection, in GFP-or E2F1-positive cells using Hoechst 33342 staining. Results are the mean ± SD of three independent experiments. (b) Western blot analysis (upper panel) and percentage of apoptotic cells, A549 and H1299 cell lines were transfected either with a control pcDNA3
, Apoptosis was assessed in GFP-or E2F1-positive cells using Hoechst 33342 staining after 6 days of geniticin selection. Results are the mean ± SD of three independent experiments. (b) Upper panel: a 96-hours cell viability assay was performed in H358/Tet-On/E2F1 or H358/Tet-On/E2F1(E132) cells co-treated with (Dox +; black symbols) or without (Dox -; white symbols) doxycyclin and increasing amounts of the FAS agonistic CH11 mAb. Results are expressed as the percentage of cell survival as compared to untreated cells. Mean value of three independent experiments ± SD performed in triplicate are presented. Lower panel: H358/Tet-On/E2F1 and H358/Tet-On/E2F1(E132) cells were treated or not with 293FasL-supernatants or 40 ng/ml TRAIL, in the presence (Dox +) or in the absence (Dox-) of doxycyclin for 72 hours. Percentage of apoptotic cells was monitored after Hoescht staining. Results are the mean ± SD of three independent experiments, H358/Tet-On/E2F1(E132) (hatched white and black bars) cells cultured in the presence (Dox +) or in the absence (Dox -) of doxycyclin for indicated times
, black symbols) or without (Dox -; white symbols) doxycyclin and increasing amounts of either etoposide or paclitaxel as indicated. Results are expressed as the percentage of cell survival as compared to untreated cells. Mean value of three independent experiments ± SD performed in triplicate are presented. (d) E2F1-deficient (E2F1 -/-, black bars) and wild-type control (E2F1 +/+ , white bars) MEFs were treated for 24 hours with increasing amounts of FasL or TRAIL and apoptosis was evaluated using Hoescht staining
, Figure 6: Downregulation of c-FLIP S is sufficient to restore tumor cells and primary E2F1-/-MEFs sensitivity to death receptor-mediated apoptosis