FIG. 5.
hr-dependent transactivation by IE1 basic domain II mutations. (A) Luciferase reporter. The luciferase gene under control of the p35 basal promoter (TATA element to the +1 RNA start site) (arrow) was linked to a single copy of the palindromic 28-mer from the hr5 enhancer. (B) Reporter transactivation. SF21 cells (2 × 106/plate) were transfected with reporter plasmid (2 μg) alone and plasmid (2 μg) encoding wild-type (wt) IE1HA or the indicated basic domain II-IE1HA mutations. Cell lysates prepared 48 h later were subjected to immunoblot analysis (1.2 × 105 cell eq) using α-HA (top panel) or assayed for luciferase activity (bottom panel). Values shown are averages ± standard deviations of triplicate transfections and are reported as RLU.