Figure EV4. Response of MDA‐MB‐231 cell lines stably transfected with shRNA targeting DAPK1, UNC5B, and NTN1 to treatments combining DAC and net1‐mAb.

• A
  Gene expression was measured by qRT–PCR after 72 h in cell lines stably transfected with shRNA. Cells were treated daily with 10 μM DAC. The level of PBGD expression was used as an internal control. Data are expressed as mean ± s.e.m. of at least 3 independent experiments. ****P < 0.0001, one‐way ANOVA.
• B–D
  Levels of DAPK1, UNC5B, and netrin‐1 were measured by immunohistochemistry in paraffin embedded xenografts of MDA‐MB‐231 stably transfected with shRNA. Left panels: levels of DAPK1 (B), UNC5B (C), and netrin‐1 (D) in xenografts of MDA‐MB‐231 shDAPK1, shUNC5B, and shNTN1, respectively. Protein levels are expressed as the percentage of total tumor surface; P‐values from at least 3 tumors per group. Middle panels: median number of apoptotic cells per mm²; values from at least 3 tumors per group. Error bars = s.e.m. Mann–Whitney test. ns = not significant. Right panels: representative sections of in vivo protein levels and induction of apoptosis in MDA‐MB‐231 tumors cells depleted of key genes from the netrin‐1 dependence receptor pathway. Scale bars = 50 μm.