Fig. 5

Targeting pertuzumab-induced signalling. a AU565 and BT474 cells were treated with pertuzumab (100 nM) for the time points indicated. Western blotting was performed with lysates from these cells with the total and phospho-antibodies indicated. Data is presented as a heat map showing the relative changes in phosphorylation (n = 3, mean); raw data and statistical analysis are presented in Additional file 2: Figure S6. b AU565 cells expressing the ERK-KTR clover biosensor were treated with pertuzumab (100 nM). Live cell imaging was performed to measure the cytoplasmic to nuclear (C/N) ratio of the ERK-KTR biosensor at the time points indicated (mean ± SEM). c Western blotting showing the co-immunoprecipitation of GRB2 and SHC with ERBB2 from BT474 cells and AU565 cells. Each line was treated with pertuzumab (100 nM) for the time period indicated and immunoprecipitation was performed with either a mouse IgG control or ERBB2 monoclonal antibody. Blots are representative of three experiments. d AU565 cells were treated with pertuzumab (100 nM) or UO126 (10 μM) for 30 min. Western blotting was performed with the antibodies indicated. Blots are representative of three experiments. e Cell viability assay was performed using the AU565 cell line with pertuzumab at the concentrations indicated, in the presence or absence of UO126 (10 μM) for 5 days (n = 6, mean ± SD, *p < 0.05). f AU565 cells were treated with the combination of pertuzumab (100 nM), BMS-777607 (1 μM) or cabozantinib (1 μM) for 30 min. Western blotting was performed with the antibodies indicated. Blots are representative of three experiments. g Cell viability assay was performed using the AU565 cell line with pertuzumab at the concentrations indicated, in the presence or absence of BMS-777607 (1 μM) or cabozantinib (1 μM) for 5 days (n = 6, mean ± SD)