Fig. 3: JMV7048 is a bona fide PXR PROTAC.
From: Targeting pregnane X receptor with a potent agonist-based PROTAC to delay colon cancer relapse
A TR-FRET competitive assay between a fluorescent agonist and JMV6945, JMV6944 or JMV7048. TR-FRET ratios (520 nm/490 nm x 104) are expressed as mean ± SD (n > 3) and normalized to DMSO (%). B Schematic representation of the competitive assays performed in (C) and JMV6945) structure. C Western-blot analysis of PXR expression in LS174T cells treated for 24 h with 5 µM JMV7048 with or without 5 µM JMV6944 or JMV6945. D Schematic representation showing the different actors in involved in the proteasome-mediated PXR degradation and their inhibitors (MLN4924 and Bortezomib, BZ). E Western blot analysis of PXR expression in LS174T cells treated 24 h with 5 µM JMV7048 or JMV7159 with or without CRBN E3 ligase inhibitor (0.5 µM MLN4924). The numbers above the bands represent PXR quantification (PXR/ACTIN vs DMSO). F Western blot analysis of PXR and ubiquitin (UBI) expression in LS174T cells lysates before (Input) and after PXR-immunoprecipitation (IP PXR). Cells were treated for 24 h with 5 µM JMV7048 in presence or absence 100 nM Bortezomib (BZ). PXR pUBI polyubiquitinated PXR. The numbers above the bands represent PXR quantification (PXR/ACTIN vs DMSO).
