Fig. 3 | Signal Transduction and Targeted Therapy

Fig. 3

From: Discovery of IHMT-337 as a potent irreversible EZH2 inhibitor targeting CDK4 transcription for malignancies

Fig. 3

IHMT-337 degrades EZH2 via CHIP-mediated ubiquitination pathway. a Effects of 24 h IHMT-337 treatment on EZH2 protein levels in both Pfeiffer and MDA-MB-231 cells. b (Left panel) Pfeiffer and MDA-MB-231 cells were treated with CHX (40 μg/ml) with or without IHMT-337 treatment (10 μM) at the indicated time points. EZH2 and GAPDH protein levels were detected by western blotting. (Right panel) The half-life of EZH2 protein was quantified and graphed. Shown are the representative results of three independent experiments. c Pfeiffer and MDA-MB-231 cells were treated with IHMT-337 and with or without the proteasome inhibitor, MG132 (5 μM) at the indicated time points, EZH2 and GAPDH protein levels were detected by western blotting. d Pfeiffer and MDA-MB-231 cells were treated with IHMT-337 for 24 h at 0, 2.5, 5, and 10 μM. IP was performed with antibodies against EZH2, ubiquitin, EZH2, and GAPDH protein levels were detected by western blotting. e Cell lysates from Pfeiffer or MDA-MB-231 cells were treated with Biotin-IHMT-337 for 4 h at 0, 1 μM. IP was performed with Streptavidin bead through streptavidin-biotin interaction, and immunoblotting was performed with antibodies against EZH2 and CHIP

Back to article page