FIG. 6.

Transfection into CNE1^Akata cells of miR200b or miR429 leads to EBV reactivation. (A) Immunoblot showing levels of the indicated proteins following transfection of CNE1^Akata cells with the indicated miRNAs. CNE1 cells latently infected with the Akata strain of EBV were transfected with 30 nM miR200a, miR200b, miR429, all three, or a negative-control miRNA. Seventy-two hours later, cells were harvested for whole-cell protein and RNA. Fifty μg of protein was loaded per well, with β-actin serving as a loading control. Relative protein levels, indicated by the numbers below the blots, were determined by densitometry, with internal normalization to β-actin and external normalization to the negative-control (Neg. cont.) sample processed in parallel. (B) Relative levels of miR200a, miR200b, and miR429 present in the cells from panel A 72 h after transfection. Levels of miR200a, miR200b, and miR429 were determined by RT-qPCR as described in Materials and Methods, with normalization to RNU38B.