Journal: Oncogene

Article Title: EZH2 PROTACs outperform catalytic inhibitors in prostate cancer by targeting a methylation-independent function of PRC2

doi: 10.1038/s41388-025-03662-z

Figure Lengend Snippet: A Scatter plot of protein expression changes based on global quantitative proteomics. C4-2B cells and 22Rv1 cells were treated with 0.1 µM of PROTAC-6272 for 6 h. Protein lysates were collected and subjected to global quantitative proteomics analysis as described in the experimental section. Data are presented as fold-change normalized to DMSO-treated cells. B LNCaP, 22Rv1, and VCaP cells were treated with DMSO, the negative control PROTAC-6286 (0.5 µM), PROTAC-6272 (0.5 µM), and EPZ-6438 (0.5 µM) for 48 h and then subjected to immunoblotting analysis. C C4-2B cells were treated with 20 μM proteasome inhibitor (MG-132) for 6 h, then treated with DMSO, negative control PROTAC-6286 (0.1 µM), and PROTAC-6272 (0.1 µM) for 24 h before immunoblotting. D C4-2B cells were treated with control (siCtrl) or siVHL for 72 h, followed by treatment with DMSO, negative control PROTAC-6286 (0.1 µM), and PROTAC-6272 (0.1 µM) for 24 h, and then subjected to immunoblotting. E C4-2B cells were transfected with HA-Ubiquitin (Ub) for 24 h, then treated with negative control PROTAC-6286 (0.1 µM) and PROTAC-6272 (0.1 µM) for 24 h and subjected to co-IP by anti-EZH2, followed by immunoblotting.

Article Snippet: For coimmunoprecipitation, C4-2B were treated with DMSO or PROTAC-6272 (0.5uM) for 3 h, and cell pellets were lysed in Co-IP buffer (50 mM tris-HCl, pH 7.4 from Life Technology, 150 mM NaCl from VWR, 1 mM EDTA from Life Technology, 1% Triton X-100 from Sigma-Aldrich) supplemented with protease inhibitor (Roche).

Techniques: Expressing, Quantitative Proteomics, Negative Control, Western Blot, Control, Transfection, Ubiquitin Proteomics, Co-Immunoprecipitation Assay