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. 2018 May 3;8:6976. doi: 10.1038/s41598-018-25404-w

Figure 4.

Figure 4

Impact of RD phosphorylation on HSP90 inhibitor effect on HSF1 activity and mobility shift in SDS-PAGE, and on HSF1 interaction with HSP90 (A) Schema of WT and dPRD FLAG-HSF1 constructs depicting RD phosphorylation sites mutated to alanine in dPRD-HSF1. (B) The N-terminal HSP90 inhibitor 17AAG enhances transcriptional activity of dPRD HSF1. HEK293 cells were transfected with hsp70-reporter construct and HSF1 C-terminal truncation constructs, treated with DMSO or 17AAG for 1 hour, heat shocked at 42 °C for 30 minutes and allowed to recover for 4 hours before harvesting. This experiment was repeated three times and values represent mean +/− S.D. (C) N-terminal HSP90 inhibitors induce a mobility shift in dPRD HSF1 similarly to that seen with WT-HSF1. HEK293 cells were transfected with WT or dPRD HSF1, treated with the three N-terminal HSP90 inhibitors as shown, and harvested and HSF1 was analyzed by WB. Full blots used to create cropped figure panels are shown in Supplementary Figure 6.