Zika virus inhibits eIF2α-dependent stress granule assembly
A. Vero cells were infected with ZIKV with an MOI of 0.5 or mock-infected and treated at 24 hpi with 500 μM Ars for 1 h to induce cellular stress. Lysates were analyzed for S51-phospho(P)-eIF2α, eIF2α (total) and NS1 by SDS-PAGE followed by Western blotting. B. Densitometry quantification of p-eIF2α was determined by ImageJ analysis. Values presented in the graph are normalized against the total amount of eIF2α in the cell lysate and represent fold change with the untreated mock-infected cells being arbitrarily set to 1. Asterisks represent the statistically significant difference between mock and ZIKV-infected cells (Two-way ANOVA; p < 0.05). C. Cellular stress was determined by IF/LSCM staining for SG markers, TIAR and phosphor-eIF2α and infected cells were identified by the presence of the NS1 viral protein. Blue arrows: uninfected cells; red arrows; infected cells. D. Quantification of the integrated density of p-eIF2α signal by ImageJ analysis.