FIG 3.

miR-21 knockdown enhances the expression of Cdc25a and viral proteins. (A) U-251MG cells were untreated (Blank) or transduced with lentiviruses expressing three different shRNAs targeting miR-21 (sh21-1, -2, and -3) or expressing a scrambled sequence control shRNA (Scram). miR-21 levels were quantified by qRT-PCR and expressed as fold differences relative to Scram. (B) miR-21 and Cdc25a mRNA levels were quantified by qRT-PCR in U-251MG cells transduced with the sh21-1 lentivirus and expressed as fold differences relative to Scram lentivirus-transduced cells (left). Cdc25a protein levels in replicate cultures were determined by Western blotting (right). Relative protein levels were determined by densitometry and are indicated below each blot. Actin serves as a loading control. (C) Transduced U-251MG cells were infected with HCMV at an MOI of 0.5, and viral and cellular proteins were detected by Western blotting at 24 and 48 hpi. The protein levels relative to Scram-transduced cells were determined by densitometry and are indicated below each blot. Actin serves as a loading control. qRT-PCR results are means ± 1 SD of data from three independent experiments, each conducted in triplicate. **, P < 0.01.