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. 2015 Jun 1;10(6):e0126645. doi: 10.1371/journal.pone.0126645

Fig 10. Puma knock-down in MEFs and its knock-out in FDMs also markedly diminish SFV-induced caspase-3 activation and apoptosis.

Fig 10

(A) Caspase-3/-7 (DEVDase) activity assay of total extracts of puromycin selected, mixed populations of SV40 TAg-transformed and 3T9-immortalized MEFs infected with lentiviruses carrying either a scrambled shRNA (sh-Ctrl) or a shRNA of mouse Puma (sh-Puma), infected with SFV for 0, 6, 14 or 24 h (hpi). As a control the data of 3T9 Puma-/- MEFs are shown. In addition, the caspase activities are compared to those from cells treated with 10 ng/ml FasL for 14 h. Data are the means of at least three independent experiments ± SEM. The p values are the following: 3T9 sh-Puma versus 3T9 sh-Ctrl: p = 0.008 for 6 h, p < 0.001 for 14 and 24 h; SV40 TAg sh-Puma versus SV40 TAg sh-Ctrl and 3T9 Puma-/- versus 3T9 WT: p < 0.001 for 6, 14 and 24 h, n = 4. (B) Annexin-V/PI FACS analysis of WT, Puma-/- and Bax/Bax-/- FDM cells infected with 10 moi of SFV for 0, 14, 24 or 36 h (hpi). Data are the means of at least three independent experiments using three different clones of WT, Puma-/- and Bax/Bak-/- cells ± SEM. The p values are the following: Puma-/- versus WT: p = 0.03 for 14 h, p < 0.001 for 24 and 36 h. Bax/Bak-/- versus WT: p = 0.01 for 14 h, p < 0.001 for 24 and 36 h, n = 4.