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. 2020 Dec 12;11(12):1061. doi: 10.1038/s41419-020-03266-3

Fig. 5. CHOP mediated tazemetostat-induced PUMA upregulation.

Fig. 5

A WT and CHOP-KO HCT116 cells were treated with 0.5 μM tazemetostat for 24 h. Indicated proteins were analyzed by western blotting. B WT and CHOP-KO DLD1 cells were treated with 0.5 μM tazemetostat for 24 h. Indicated proteins were analyzed by western blotting. C WT and CHOP-KO HCT116 cells were treated with the combination of 2.5 μg/mL 5-FU and 0.5 μM tazemetostat for 24 h. Apoptosis was analyzed by flow cytometry. D WT and CHOP-KO DLD1 cells were treated with the combination of 2.5 μg/mL 5-FU and 0.5 μM tazemetostat for 24 h. Apoptosis was analyzed by flow cytometry. E Chromatin immunoprecipitation (ChIP) was performed using anti-CHOP antibody on HCT116 cells following tazemetostat (0.5 μM) treatment for 12 h. The IgG was used to control for antibody specificity. PCR was carried out using primers surrounding the CHOP binding sites in the PUMA promoter. The results were expressed as the mean ± SD of three independent experiments. **P < 0.01.