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. 2017 Feb 17;114(10):2681–2686. doi: 10.1073/pnas.1621508114

Fig. 6.

Fig. 6.

BAX- and BAK1-dependent generation of mitochondrial reactive oxygen species causes cytoplasmic accumulation of TIPARP to limit SINV replication. (A) Bax+/+/Bak1+/+ and Bax−/−/Bak1−/− MEFs stably expressing TIPARP-FLAG were infected with SINV (MOI = 5) or stimulated with CCCP (10 μM) for 12 h. Fixed samples were subjected to immunocytochemistry analysis of TIPARP-FLAG and Hoechst 33342 staining of genomic DNA. (B) Bax+/+/Bak1+/+ and Bax−/−/Bak1−/− MEFs were infected with SINV (MOI = 5) or stimulated with CCCP (10 μM) for 12 h, and then stained with MitoSOX Red. Samples were subjected to flow cytometric analysis to measure levels of mitochondrial reactive oxygen species. (C and D) Bax+/+/Bak1+/+ and Bax−/−/Bak1−/− MEFs (C) or primary Mavs+/− and Mavs−/− MEFs (D) stably expressing TIPARP-FLAG were then infected with SINV (MOI = 5) for 24 h. Viral titers in culture supernatants were determined by 50% tissue culture infectious dose assay. (Scale bar, 20 μm.) Experiments were performed three times, and representative data are shown (means ± SD of three independent samples). *P < 0.05.