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. 2018 Jun 19;32(12):6883–6897. doi: 10.1096/fj.201800413RR

Figure 6.

Figure 6

Activated JNK enhances c-Jun phosphorylation and promotes caspase 3 cleavage. A) MEFs were transfected with either FLAG-MKK7-JNK1 (JNK active) or FLAG-MKK7-JNK1 APF variant (JNK inactive) for 24 h before exposure to C6-Cer (Cer6, 60 μM) for 10 h. B) Quantification of phosphorylation of c-Jun (Ser63) and cleaved caspase 3 levels. *P < 0.05, ***P < 0.01. C) R28 cells were transfected with FLAG-MKK7-JNK1 (JNK active) or FLAG-MKK7-JNK1 APF variant (JNK inactive) for 16 h before exposure to 60 μM Cer6 or 1.2 μl/ml DMSO vehicle (Veh) for 8 h. D, E) Quantification of phosphorylation of c-Jun (Ser63) (D) and cleaved caspase 3 (E) levels. FLAG-MKK7-JNK1 was observed at 102 kDa. Bands are from the same blot, but are not contiguous (A). Protein loading was assessed by protein staining (Protein S). Protein molecular mass (kDa) is indicated at the right of the blots. Blots shown are representative of results for 2 experiments; within each experiment, 3 independent samples were analyzed. Results are means ± sem (n = 3). Different letters (a–c) above the scatter plots (D, E) denote significantly different results. P < 0.05.