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

Fig. 5. Oncogenic HRAS induction causes a tension differential in the MCF10A bilayers.

Fig. 5.

Representative confocal images of nontransformed monolayer (A) and HRAS-transformed bilayer (D) at t = 24 hours from oncogene induction stained for pMLC2, F-actin, and DAPI (nuclei, shades of grays). Focal planes are orthogonal to the substrate matrix. Scale bar, 20 μm. (B, E, and G) Representative confocal images of entire nontransformed monolayer (B) and HRAS-transformed bilayer [(E) bottom layer and (G) top layer of the bilayer]. Focal plane crosses the tissue parallel to the substrate matrix. pMLC2, F-actin, and DAPI (nuclei) are labeled either in color code (red, gray, and blue, respectively) or shades of grays. Scale bars, 50 μm. (C) Relative fold change in averaged global intensity relative to center of the monolayer of pMLC2 fluorescence from edge to edge of the circular epithelial domain of nontransformed (blue) and HRAS-transformed (pink) monolayers (n = 3 and means ± S.D. in blue). (F) Relative fold change in averaged global intensity relative to center of the monolayer of F-actin fluorescence from edge to edge of the circular epithelial domain of nontransformed (blue) and transformed (pink) monolayers (n = 3 and mean ± S.D. in blue) and (H) individual averaged intensity profiles of pMLC2 (red) and F-actin (gray) of a representative section of HRAS-transformed top layer.