Figure 7.

Activation of PI3K/Akt signaling pathway suppresses EV71-induced apoptosis in RD and HEK 293 cells. (A) RD cells were exposed to EV71 infection at an MOI of 2. After 12 h, the cells were pre-incubated for 30 min in the presence of NucView488 (5 μM). Fluorescence micrographs were taken at 12 hpi [representative images are shown; at least 5 individual cells were analyzed in each independent experiment (n = 3)]. (B,C) RD and HEK293 cells were infected with EV71 at an MOI of 2. At the indicated time points post-infection, the percentage of apoptotic cells was measured by Nucview 488 (Biotium) described in section Materials and Methods. (D,E) RD and HEK293 cells were transfected with or without hsa-miR-494-3p mimics or inhibitor for 48 h, or pre-incubated with or without Wortmannin (100 nM) for 1 h, and then followed by infection with EV71 for 6 and 12 h at an MOI of 2. Nucview 488 assay (flow cytometry) was employed to measure the percentage of apoptotic cells at the indicated time points. The experiments were repeated three times (p < 0.01 by ANOVA).