Fig. 4.

Mt-YAP5SA expression in BRCA1-deficient mammary epithelial cells confers EGF-independent proliferation and tumorigenesis. (A–C) HEK293A or MCF10A cells were infected with lentivirus encoding shBRCA1 and indicated viruses. Then after 48 h, cells were seeded in 96-well plates, and treated with wst1 reagent to compare cell growth as described in Materials and Methods. (D) MCF10A cells were infected as described in A and then grown on matrigel in presence or absence of EGF. Fresh medium lacking EGF was added every 4 d. (E and F) Representative immunohistochemistry pictures and relative tumor volume. (G) MCF10A cells were treated with hydroxyurea as described in Materials and Methods, washed, and then incubated in six-well plates. Then after 10–12 d, cells were fixed, stained with crystal violet, and cell survival was determined by measuring absorbance at 590 nM. Results are expressed as means ± SEMs. Only values of P < 0.05 were considered significant (****P < 0.0001), by ANOVA test.