Fig. 3: PUMA is required for the apoptotic activity of MEK inhibitors. | Oncogenesis

Fig. 3: PUMA is required for the apoptotic activity of MEK inhibitors.

From: RETRACTED ARTICLE: MEK inhibitors induce apoptosis via FoxO3a-dependent PUMA induction in colorectal cancer cells

Fig. 3

a Parental, p53-KD and PUMA-KD RKO cells were treated with 0.1 μmol/L trametinib or 0.1 μmol/L selumetinib for 24 h. Apoptosis was analyzed by a nuclear fragmentation assay. b Parental and PUMA-KD RKO cells were treated with 0.1 μmol/L trametinib or 0.1 μmol/L selumetinib for 24 h. Apoptosis was analyzed by Annexin V/PI staining followed by flow cytometry. c Parental and PUMA-KD RKO cells were treated with 0.1 μmol/L trametinib or 0.1 μmol/L selumetinib for 24 h. Caspase 3/7 activity was determined by fluorogenic analysis. d Parental and PUMA-KD RKO cells were treated with 0.1 μmol/L trametinib for 24 h. Cleaved caspase 3 and 9 were analyzed by Western blotting. e The cytoplasm and mitochondria were fractionated from parental and PUMA-KD RKO cells treated with 0.1 μmol/L trametinib for 24 h. The distribution of cytochrome c was analyzed by Western blotting. β-Actin and cytochrome oxidase subunit IV (Cox IV) were analyzed as the control for loading and fractionation. f Parental and PUMA-KD RKO cells were treated with 0.1 μmol/L trametinib for 24 h. Colony formation assay was performed by seeding an equal number of treated cells in 12-well plates and then staining attached cells with crystal violet 10 days later. Left, representative pictures of colonies; Right, quantification of colony numbers. The results in a, b, c and f are expressed as the means ± SD of three independent experiments. **P < 0.01, *P < 0.05

Back to article page