Fig. 2
9F7-F11 induces c-FLIP downregulation by proteasomal degradation. a Cancer cells were incubated with NRG1 or 9F7-F11. After cell lysis at different time points, c-FLIPL, USP8 and USP9X expression, as well as ITCH expression and phosphorylation (p) were analyzed by western blotting. b BxPC3 cells were incubated with 9F7-F11 for 3 h. HER3, c-FLIPL and c-FLIPS expression were analyzed by western blotting. c BxPC3 and MDA-MB-468 cells were incubated with NRG1 or 9F7-F11 for 48 h or 96 h, before detection of HER3 and c-FLIPL expression by western blotting. d After pre-incubation or not with 10 μM MG132 for 4 h, BxPC3 cells were incubated with 9F7-F11 with or without NRG1. After cell lysis, expression of HER3 and c-FLIPL was assessed in whole cell lysates by western blotting. The rabbit anti-HER3 polyclonal antibody C-17 (Santa Cruz Biotechnology) was used for detection. Protein level was measured with the ImageJ software and indicated as signal intensity (SI), relative to untreated control (SI = 1.0 ± .0). Significant increase or decrease of the densitometry, compared to control, is indicated in bold. β-tubulin was evaluated as loading control