Localization of PCBP1 in ER-derived membrane-associated complex during EV71 replication.

(A) RD cells were mock-infected or infected with EV71 at an MOI of 5 for 8 h. Cells were probed with rabbit anti-PCBP1 antibody and mouse anti-dsRNA antibody, stained with ER-Tracker, observed by confocal microscopy. PCBP1 (red), dsRNA (green), ER-Tracker (blue). Bar = 20 µm. (B) Membrane flotation analysis of location of PCBP1. RD cells were infected with or without EV71 at an MOI of 5 for 8 h. Cell lysates from mock-infected and infected cells treated with or without Triton X-100 were separated by sucrose gradient sedimentation. Each fraction was blotted with specific antibodies to the ER marker protein (calnexin, CNX), PCBP1, and EV71 2C, respectively. Fractions 1 to 3 represent the membrane fraction. Fractions 4 to 9 represent the cytosolic fraction.