Figure 3
Trastuzumab and EV20 cooperate to disrupt the NRG-1β/ErbB-3/Akt axis and inhibit cell proliferation. (a) Serum-starved (24 h) cells were treated with EV20, trastuzumab or their combination (10 μg/ml of each antibody) for 2 h and then stimulated with NRG-1β (10 ng/ml) for 5 min; cell lysates were blotted as indicated. (b) For anchorage-independent growth assay, cells were treated twice a week with EV20, trastuzumab or their combination for 3 weeks. Data are represented as number of colonies per field. The values are expressed as mean±s.d. of triplicate experiments. *P<0.05; **P<0.01. (c) BxPC-3 cells were grown under the condition indicated and treated with the antibodies or PBS (control) for 72 h before performing MTT staining. Results are expressed as mean of quadruplicate experiments. HPAF-II cells were grown in medium containing 0,5% fetal bovine serum and treated with the antibodies or PBS (control) for 96 h before performing MTT assay. Medium was renewed every 48 h. Results are expressed as mean of triplicate experiments. *P<0.05; **P<0.01.
