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. 2017 Nov 6;127(12):4449–4461. doi: 10.1172/JCI96324

Figure 5. Stabilization of c-FLIPL by Nrg4 attenuates stress-induced cell death in hepatoma cells.

Figure 5

The following experiments were performed in Hepa 1 cells stably expressing ErbB4. Cells were cultured in serum-free medium during treatment. (A) Immunoblots of total lysates from cells treated with PA/TNF-α in the absence or presence of 100 ng/ml Nrg4 for 6 hours. (B) Immunoblots of total lysates from cells treated with PA/TNF-α without or with 100 ng/ml Nrg4 and chased for different times in the presence of 2 μM CHX. (C) Immunoblots of total lysates from cells treated with PA/TNF-α without or with 100 ng/ml Nrg4 and chased for 6 hours in the presence of 10 μM MG132. (D) Immunoblots of total lysates from treated cells. Hepa 1 cells stably expressing ErbB4 were transduced with Ad-GFP or Ad–c-FLIPL adenoviral vector. Transduced cells were treated with 100 μM PA for 2 hours followed by addition of 20 ng/ml TNF-α and 100 ng/ml Nrg4 for 20 hours. (E) LDH release by treated Hepa 1 cells (20 hours treatment). Data represent mean ± SEM. **P < 0.01, 1-way ANOVA. (F) Flow cytometry analysis (20 hours treatment). Data represent mean ± SEM. **P < 0.01; ***P < 0.001, 1-way ANOVA.