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. 2010 Jul 28;84(19):10329–10343. doi: 10.1128/JVI.00923-10

FIG. 8.

FIG. 8.

miRNA-mediated EBV reactivation is mediated via the ZEB-binding element of Zp. (A and C) Immunoblots showing levels of the indicated proteins following transfection of 293B95.8-WT and 293B95.8-ZVmt cells with the indicated miRNAs. 293 cells latently infected with a bacmid containing the B95.8 strain of EBV (A) or a variant of this bacmid with a 2-bp substitution mutation in the ZV element of Zp (C), respectively, were transfected with 30 nM concentrations of the indicated miRNAs. Forty-eight hours later, 20 ng/ml TPA and 3 mM sodium butyrate were added to the medium, and incubation was continued for another 48 h. The cells were then harvested for whole-cell protein and RNA. Thirty μg of protein was loaded per well for analysis of ZEB1 and ZEB2, and 10 μg of protein was loaded per well for analysis of BZLF1 and BMRF1. (B and D) Relative levels (Rel. Amt.) of miR200a, miR200b, and miR429 present in the cells from panels A and C, respectively. Levels of miR200a, miR200b, and miR429 were determined by RT-qPCR as described in Materials and Methods, with normalization to RNU38B.