Fig. 6.

Elevated HSP70 enhances the expression of Ad5 proteins when PML-II is reduced. (a, e) Samples were harvested from HelaEV and HelaΔII cells and analysed for HSP70 mRNA (a) or a selection of HSP mRNAs (e) by RT-qPCR, or (a, lower) for HSP70 protein by Western blot. (b) Hela cells were transfected or not with 125 pmol ml⁻¹ siRNA as indicated and RNA was harvested after 48 h for analysis of HSP70 mRNA by RT-qPCR. (c) HelaEV and HelaΔII cells were transfected with 125 pmol ml⁻¹ HSP70 or control siRNA for 48 h, then infected with Ad5 wt300 for 20 h before RNA was harvested and analysed for hexon mRNA by RT-qPCR. (d) HelaΔII cells were treated with siRNA and infected as in (c), then lysed and analysed by Western blotting as in Fig. 5. In (a–c), data were standardized to an internal control and then normalized to values from: (a) HelaEV; (b) siControl-treated Hela; (c) siControl-treated HelaEV. Panel (e) shows mRNA amounts measured separately for each amplicon, standardized in each case to an internal control. Graphs show the means and standard deviation of three technical replicates.