Fig. 1: HRO761 is a specific and potent WRN inhibitor.
From: WRN inhibition leads to its chromatin-associated degradation via the PIAS4-RNF4-p97/VCP axis
a Chemical structure of HRO761 (WRNi). b WRN inhibition leads to induction of the DNA damage response in MSI-H cells. Phospho-histone H2A.X (Ser139) (γH2A.X) staining was used to visualize DNA damage in MSI-H (HCT-116, SW-48) and MSS (HT-29, U2OS) cells after 24 h treatment with indicated compounds. Scale bar = 20 μm. c Quantification of γH2A.X signal levels in cells in (b), normalized to DMSO and Etoposide. Graphs represent averages of n = 3 plate replicates. d WRN inhibition by WRNi leads to an induction of the DNA damage response resulting in apoptosis. HCT-116 cells were treated with 10 μM WRNi and analyzed by Western Blot (WB) to assess DNA damage response markers. GAPDH was used as a loading control. e Dose-response curves are measuring the viability of HCT-116 cells or HT-29 cells after treatment with WRNi for 4 days. Graphs represent averages from n = 6 plates. f Dose-response curves measuring in vitro WRN or BLM unwinding activity after treatment with WRNi. Graphs represent averages from n = 4 replicates. WRN and BLM unwinding activity is normalized to DMSO. g Summary of IC50 or EC50 values of WRNi in the indicated biochemical and cellular assays. All curve fits were done by fitting a 4-parameter logarithmic regression curve. All error bars represent standard deviation (s.d.). DMSO is dimethyl sulfoxide; BLMi is BLM inhibitor Compound 2; Etopo is etoposide. MW is molecular weight. Source data are provided as a Source Data file.
