FIG 3.

WC5 abrogates the IE2-mediated repression of MIEP. (A) U373-MG cells were transfected with a luciferase reporter plasmid containing HCMV MIEP (pMIEP-crs) with either wild-type (wt) or mutated (mut) crs or cotransfected with an IE2-expressing plasmid (pSGIE86) and then treated with 0.2% DMSO or 50 μM WC5 or WC5E. (B) U373-MG cells were cotransfected with the MIEP-crs wt plasmid and plasmids expressing mutant P535A/Y537A and H446A/H452A IE2 proteins and then treated with 0.2% DMSO or 50 μM WC5 or WC5E. (A and B) Transfection mixtures also contained a plasmid constitutively expressing Renilla luciferase to normalize variations in transfection efficiency. The reported values represent the means ± SD from three independent experiments in duplicate and are expressed as relative luciferase units (RLU) (i.e., light units of firefly luciferase per 10⁴ Renilla luciferase light units as determined at 48 h posttransfection); *, P < 0.05, and ***, P < 0.001, versus calibrator sample (pMIEP crs wt or mut plus pSGIE86 wt or mut plus DMSO).