Figure 2.

GSDMD is responsible for ZIKV-induced cytotoxicity in human glioblastoma cells

(A) A549 cells with or without ZIKV infection were transfected with V5-tagged GSDMD or GSDME. Cell lysates were examined by western blot analysis. White arrows, full-length GSDMD or GSDME; black arrow, cleaved product. (B) GSDMD genomic DNA sequencing of SF268 wild-type (WT) and knockout (GSDMD^−/−) cells. The single guide RNA sequence designed for CRISPR-Cas9 genome editing is marked in a black frame. (C–E) WT and GSDMD^−/− SF268 cells were infected with ZIKV for western blot analysis (C, bottom panel), LDH release (C, top panel), IL-1β secretion (D), and phase-contrast imaging (E). Data are mean ± SD (n = 3 per group). ∗∗∗p < 0.001. (F) WT (stably RFP-labeled) and GSDMD^−/− (stably GFP-labeled) SF268 cells were cocultured overnight and synchronize infected with ZIKV for another 24 h, then were examined by fluorescent and phase-contrast microscopy. Yellow arrows, damaged cells.