Activation of PI3K, Akt, and ERK during early rotavirus infection leads to V-ATPase-dependent endosomal acidification required for uncoating
Fig 5
Involvement of PI3K/Akt and MEK/ERK signaling pathways in RVA uncoating.
(A-D) MA104 cells were pretreated with or without wortmannin or U0126 for 1 h at 37°C and then infected with the DS-1 or NCDV strains (MOI = 10 FFU/cell) for the indicated time. After fixation and permeabilization, the cells were prepared for confocal microscopy using anti-VP8* antibody, anti-EEA1 antibody (A and B), and anti-LAMP2 antibody (C and D), and the relevant secondary antibodies. Actin cytoskeleton was stained with AF488-labeled phalloidin. (E and F) MA104 cells were transfected with scrambled siRNA or siRNAs specific for PI3K p85α or MEK, and then infected with the DS-1 and NCDV strains (MOI = 10 FFU/cell). After fixation and permeabilization, the cells were prepared for confocal microscopy using anti-VP8* antibody, anti-EEA1 antibody (E), and anti-LAMP2 antibody (F), and the relevant secondary antibodies. Representative images are shown. The scale bars correspond to 5 μm.