FIG 7.

miR-21 is downregulated by HCMV proteins IE1, pp71, and UL26. (A) U-251MG cells were transduced with an empty vector control lentivirus (Ctrl) or lentiviruses expressing IE1, IE2, pp65, pp71, or UL26. The levels of each protein were determined by Western blotting. Actin serves as a loading control. (B) miR-21 levels in transduced cells were quantified by qRT-PCR and expressed as fold differences relative to Ctrl-transduced cells. (C) HEK293T cells were cotransfected with a reporter plasmid encoding luciferase under the control of the miR-21 promoter with plasmids expressing IE1, IE2, pp71, or UL26. The luciferase activities were measured at 48 h posttransfection and are expressed as fold differences relative to IE2-transfected cells. (D) U-251MG cells were mock treated (M) or exposed to UV-inactivated HCMV equivalent to an MOI of 10 (UV). miR-21 levels were determined by qRT-PCR immediately (0 h) or after 2 h of exposure and expressed as fold differences relative to mock-treated cells. Luciferase and qRT-PCR results are means ± 1 SD of data from three independent experiments, each conducted in triplicate. *, P < 0.05; **, P < 0.01.