Fig. 5.

Mutant HRAS^Q61R expression induces transformation and growth in non-malignant breast epithelial cells. a Representative images of soft agar anchorage-independent growth assay of parental MCF-10A PIK3CA-wild-type (MCF-10A^P), MCF-10A PIK3CA H1047R-mutant (MCF-10A^H1047R), and MCF-12A cells stably expressing empty vector (EV), HRAS-wild-type (HRAS^WT), or HRAS Q61R-mutant (HRAS^Q61R) protein (scale bars, 2 mm). Boxplots showing the quantification of the size of colonies (see Methods). The mean value of the size of colonies, and the 75th and 25th percentiles are displayed at the top and bottom of the boxes, respectively. b MTT cell proliferation assay of MCF-10A^P, MCF-10A^H1047R, and MCF-12A cells stably expressing EV (black), HRAS^WT (yellow), or mutant HRAS^Q61R (red) protein. c The migratory effects of MCF-10A^P, MCF-10A^H1047R, and MCF-12A cells stably expressing EV, HRAS^WT, or mutant HRAS^Q61R were analyzed using the wound-healing assay at 0 and 24 h. Scale bars, 500 µm. In a−c, data are representative of three independent experiments. Error bars, s.d. of mean (n = 3). n.s. = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; two-tailed unpaired t-tests