Figure 3. CstF-64 in EV71-infected cells.

(A) After RD cells have been infected with EV71 (m.o.i. = 40), 3C^pro and CstF-64 protein in total cellular proteins of infected RD cells (+) or mock infected cells (-) were detected at various hours post-infection (h.p.i.) using specific antibodies. A potential cleavage product of CstF-64 was also indicated (*). The detection of β-actin was used as a loading control (B) RD cells with transient FLAG-CstF-64 overexpression were also infected with EV71 at an m.o.i. of 40 and the total cellular proteins from mock infected (-) and infected cells (+) were harvested at 6 and 8 h.p.i.. FLAG-CstF-64 was detected using FLAG-specific antibody. The other cleaved FLAG-peptides were indicated as CP1 and CP2 (C) Cytoplasmic (C) and nuclear (N) fractions from EV71-infected RD cells at 8 h.p.i. were extracted and CstF-64 or 3C^pro in each of the fractions were detected using specific antibodies. Detections of β-actin and histon deacetylase (HDAC) were used as cytoplasmic and nucleus protein controls. (D) The locations of CstF-64 in uninfected (Mock) or EV71-infected cells at 2, 4, 6, 8 and 10 h.p.i. were detected using specific antibody. The detection of viral 2B protein was applied as an infection-positive marker. The nuclei of cells were stained using Hoechst dye.