Fig. 5: DCAF1 and EZH2 inhibitors as potent modulators of cell growth and target gene expression.

a EZH2-depleted SW620 cells were complemented with lentiviruses expressing 3×FLAG-EZH2 wild type, T367A phospho-blocking mutant, or T367D phospho-mimicking mutant. The cells were then treated with DMSO or B32B3 (0.9 µM) for 3 days, and WST assays were performed to measure their viability. Data are represented as mean ± SEM of three independent experiments. P values were calculated using two-way ANOVA with post-hoc Tukey’s test for multiple comparisons. *P < 0.05, **P < 0.01 and ***P < 0.001 versus Ctrl sh. b After treating SW620 cells as in (a), total RNA was prepared and analyzed by RT-qPCR using primers listed in Supplementary Table 4. Data are represented as mean ± SEM of three independent experiments. P values were calculated using two-way ANOVA with post-hoc Tukey’s test for multiple comparisons. **P < 0.01 and ***P < 0.001 versus Ctrl sh. c After treating SW620 cells as in (a), whole cell lysates were prepared and subjected to Western blot analysis with the indicated antibodies. Data are representative of three independent experiments. d SW620 colon cancer cells were treated with increasing concentrations of Taz (0–10 µM) in the absence or presence of B32B3 (0.9 µM) for 72 h, and their viability was measured by WST assays. Data are represented as mean ± SEM of three independent experiments. P values were calculated using two-way ANOVA with post-hoc Tukey’s test for multiple comparisons (***P < 0.001). (See also Supplementary Fig. 13.). e SW620 cells were treated with IC50 concentrations of B32B3 (0.9 µM) and/or Taz (15 µM) for 72 h, and total RNA was prepared and analyzed by RT-qPCR using primers listed in Supplementary Table 4. Data are represented as mean ± SEM (n = 3 biologically independent experiments). P values were calculated using two-way ANOVA with post-hoc Tukey’s test for multiple comparisons. ***P < 0.001 versus DMSO; ###P < 0.001 versus B32B3. f SW620 cells were treated as in (e), and cell lysates were prepared and analyzed by Western blotting with the antibodies indicated on the left. Data are representative of three independent experiments. Source data are provided as a Source Data file.