(A) RD cells were transfected with control siRNA or siRNAs targeting PI4KB, ARF1, and GBF1. At 36 h post-transfection, cells were mock-infected (−) or infected (+) with EV71 (MOI = 10) for 12 h. The cell lysates were then analyzed by western blotting with the indicated antibodies. (B) RD cells were infected with EV71 (MOI = 10) for 12 h; the cells were then fixed and stained with mouse anti-p97 and rabbit anti-PI4KB/ARF1/GBF1 antibodies (nuclei, blue; p97, green; PI4KB/ARF1/GBF1, red). (C) RD cells were treated as described in (B) and then stained with mouse anti-2C and rabbit anti-PI4KB/ARF1/GBF1 antibodies. (D) RD cells stably expressing p97-GFP were infected EV71 (MOI = 10) for 12 h, fixed, and stained with rabbit anti-PI4KB and mouse anti-2C antibodies (nuclei, blue; p97, green; PI4KB, red; 2C, purple). The upper panels show an image of each channel and the lower panels show merged pictures of two or four channels as indicated. (E) Cells were treated as described in (D), except that RD cells were stably expressing ARF1-GFP and stained with rabbit anti-PI4KB and mouse anti-p97 antibodies (nuclei, blue; ARF1, green; PI4KB, red; p97, purple).