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. 2017 Jul 3;36(17):2510–2528. doi: 10.15252/embj.201696089

Figure EV4. Skp2 inactivation effectively suppreses YAP‐driven aberrant stiff 3D matrix‐evoked epithelial tissue behaviors.

Figure EV4

  1. MCF10A cells were embedded in soft or stiff 3D BM/COL1 gels. After 8 days of growth, the cells were fixed and stained with phalloidin and a YAP‐specific antibody. DAPI was used as a nuclear counterstain. Images were taken by confocal microscopy. Scale bars: 50 μm.
  2. Relative mRNA levels for the indicated genes in MCF10A cells treated as in (A), as measured by qPCR. Error bars indicate standard error of the mean (s.e.m.) for four independent experiments. Statistical significance, as determined by a two‐tailed t‐test, is indicated in the graph. N.S. indicates non‐significance.
  3. Vector or 5SA‐YAP‐expressing MCF10A cells infected by the indicated shRNA lentiviruses were embedded in soft or stiff 3D BM/COL1 gels. After 7–8 days of growth, the cells were imaged by phase contrast (gray) or confocal microscopy (blue: DAPI). Scale bars: black, 100 μm; white, 50 μm.
  4. Empty vector‐ or 5SA‐YAP‐expressing MCF10A cells were embedded in stiff 3D BM/COL1 gels. After 7–8 days of growth, the cells were treated as in (A). Scale bars: 50 μm.
  5. Vector‐ or 5SA‐YAP‐expressing MCF10A cells were embedded with either DMSO or the Skp2 inhibitor SZL P1‐41 (20 μM) in soft or stiff 3D BM/COL1 gels. After 7–8 days of growth, the cells were fixed and imaged by phase contrast microscopy. Images are representative of three independent experiments. Scale bars: 100 μm.