Fig. 7: Investigating the mechanisms of HIF-1α regulation by dFLASH-HIF inhibitor hit RQ500235 and activator hit RQ200674. | Nature Communications

Fig. 7: Investigating the mechanisms of HIF-1α regulation by dFLASH-HIF inhibitor hit RQ500235 and activator hit RQ200674.

From: dFLASH; dual FLuorescent transcription factor activity sensor for histone integrated live-cell reporting and high-content screening

Fig. 7

a, b Inhibitor RQ500235 identified from the bimodal screen (a) represses DMOG induced Tomato in dFLASH-HIF cells in a dose dependent manner (n = 2 biological replicates) and (b) decreases expression of HIF-1α protein as assessed by immunoblot of whole cell extracts from endogenous HA-Flag tagged HIF-1α in HEK293T cells. S.E. short exposure; L.E. long exposure. Representative of 3 independent replicates. c RT-qPCR shows HIF-1α transcript is significantly decreased in HEK293T cells treated for 6 h with RQ500235 (n = 3 biological replicates, *p = 0.0139 assessed by a two-tailed t-test with Welch’s correction, presented as mean relative expression ± SD). d Activator RQ200674 identified from the bimodal screen recapitulated activation of dFLASH-HIF at 50 μM in HEK293T cells (n = 2 biological replicates). e In vitro iron chelation assay of RQ200674 which displays weak chelating activity at 236 μM from line of best fit (n = 3 independent experiments) compared to positive control iron chelator and HIF-1α activator, dipyridyl. Source data are provided as a Source Data file.

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