Fig. 4

ZIKV-LAV infection induces apoptotic and necrotic cell death in human GBM cells. A–B Caspase-3 activation for apoptosis in virus-infected A DBTRG and B T98G cells detected by luminescence assay on days 1 and 3 post-infection. C Western blot detection of cleaved caspase-3 in infected cell lysates at day 2 post-infection. D Lactose Dehydrogenase (LDH) release by necrotic cells at day 3 post-infection. E Gating strategy for the simultaneous detection of apoptotic and necrotic DBTRG cells by flow cytometry. F Representative scatter dot-plots of DBTRG cells during early (day 1) and late (day 3) stages of infection with ZIKV-LAV. Cells were co-stained and detected with apopxin-FITC and 7-AAD-mKATE. G–I Quantification of the fraction of G apoptotic, H late apoptotic/necrotic, and I necrotic DBTRG cells by flow cytometry during late stage of infection with ZIKV-LAV. Values are presented as % of live cells. Data are presented as mean ± SEM. Non-parametric Mann–Whitney test or Kruskal–Wallis test with Dunn’s post-hoc correction was used to compare groups. p-values are shown accordingly: *p < 0.05. **p < 0.005, ***p < 0.001