Skip to main content
. 2021 Oct 13;7(42):eabg6467. doi: 10.1126/sciadv.abg6467

Fig. 1. Morphological characterization of normal and HRAS-transformed MCF10A tissues.

Fig. 1.

(A) Schematic of the experimental setup. (B) HRAS induction confirmed by increase in phosphorylation of mitogen-activated protein kinase (MAPK) (ERK1/2) shown in representative Western blots. (C) Phase contrast time-lapse of selected nontransformed (control) and HRAS-transformed MCF10A monolayers (imaging starts at t = −4 hours, and HRAS activation is induced at t = 0 hours; scale bars, 200 μm). (D) Contours of epithelia shown in (C) (time progresses with blue, red, green, and yellow colors). (E) Time evolution of the surface area of the epithelia’s footprint on the substate matrix (epithelial domain). (F) Time evolution of the major and minor diameters of the epithelia domain in the case of nontransformed and HRAS-transformed epithelia. (G) Cell number in HRAS-transformed epithelia at 12 hours after induction [start of area decline, (E)] compared to nontransformed epithelia at 12, 24, and 44 hours. Kruskal-Wallis test + Dunn’s multiple comparisons test. (H) Time evolution of the epithelial domain’s aspect ratio (i.e. major axis divided by minor axis of the epithelial domain). (I) Time evolution of the epithelial domain’s circularity. (E to I) Statistics over 15 nontransformed epithelia and 16 HRAS-transformed epithelia from at least four independent experiment repeats. Each epithelium was imaged for at least 50 hours. (J) Time evolution of the surface area of the nontransformed (n = 7) and HRAS-transformed (n = 8) epithelia domains during cell cycle arrest (9 μM RO-3306). (E, F, and H to J) Time evolution graphs represent means ± SEM of medians at each time point. Two-way analysis of variance (ANOVA) with Bonferroni posttest, *P < 0.05. a.u., arbitrary units.