Fig. 2: ECM adhesion and TGFβ signalling regulate cell invasion in OHGs.

a pFAK, pMLC, YAP/TAZ (green; left to right) and nuclei (blue) staining of OVCAR8 cells in collagen or organo-hydrogel (OHG) after 48 h in culture (representative images from n = 3 gels). b Quantification of immunofluorescence signal intensity of cytoplasmic pFAK, pMLC, relative to the average in collagen and cytoplasmic:nuclear ratio of YAP/TAZ in OVCAR8 cells embedded in collagen or OHG for 48 h (n = 24 cells). c F-actin (yellow), nucleus (blue) and BODIPY (grey) staining of an OVCAR8 cell spread at the ECM-microdroplet interface. d Shape descriptors of OVCAR8 nuclei after 24 h in collagen or OHG (n = 53 nuclei). e Relative spheroid area of OVCAR8 after 7 d in OHG (n = 10 spheroids). f BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells (49 μm into the tissue from the surface) after 7 d in culture (representative images from n = 3 experiments). g Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d (n = 3 tissues from distinct donors). Rhombuses indicate the average. h Nuclei (red) of OVCAR8 cells in OHGs after 7 d treatment (representative images from n = 14 spheroids). i Quantification of spheroid area relative to DMSO control of OVCAR8 cells in OHGs (n = 14 spheroids). i F-actin (green) and nuclei staining of CAOV3 spheroids embedded in OHGs for 7 d (representative images from n = 6 spheroids). k Quantification of spheroid area in CAOV3 cells in OHGs after 7 d (n = 6 spheroids). l F-actin (green) and nuclei (blue) staining of Kuramochi spheroids in collagen or OHG for 7 d with or without TGFβ (representative images from n = 3 spheroids). For the data in (b, d, g, i and k), a two-sided unpaired t test was performed. For data in (e), a one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed. Scale bars, 25 μm (a, c), 100 μm (f, I), 200 μm (h, j). Source data are provided as a Source Data file.