Fig. 5: Experimental test of the predicted morphometric signatures of tumor cells.
From: Mechanical control of cell fate decisions in the skin epidermis
a Immunostaining for β4-integrin, YFP and SmoM2 in Krt14-CREER/YFP and Krt14-CREER/SmoM2 epidermis at different time points upon tamoxifen administration. Scale bars: 10 μm. b Quantification of the basal clone size in clones from Krt14-CREER/YFP and Krt14-CREER/SmoM2 at different time points upon tamoxifen administration (n = number of clones quantified from 4 animals). Error bars represent the mean ± SEM. c Schematic depicts the quantity that is used to measure tension homogeneity between adjacent clones-- contact angle (θc). For more details see Section: “Methods”. Using representative z-stack images of mouse skin (IFE), showing Phalloidin (gray), DAPI (blue) for nuclei, immunostaining for β4-integrin (red) for BM and SmoM2 (green), an example for contact angle measurement is shown for a 3-cell clone. More details in Section: “Methods”. Scale bars: 10 μm. Contact angle distributions (d) and averages (e) are shown for week 1 (top in (d), from 3 animals each in (d) and (e)) and week 2 (bottom in (d), from 2 animals each in (d) and (e)). Dotted gray line is drawn at 90∘ for reference, with smaller angles found on average for Krt14-CREER/SmoM2 cells. Error bars represent the mean ± SEM. f Basal contact area of individual cells, normalized by cellular volumes, shown for Krt14-CREER/YFP and Krt14-CREER/SmoM2 clones (n = number of clones quantified from 3 animals). Error bars represent the mean ± SEM. g Representative images of mouse skin IFE are shown for Krt14-CREER/YFP and Krt14-CREER/SmoM2 clones showing the 3D reconstruction of the BM (β4-integrin) in brown and clones in green, for times—1 and 2 weeks after tamoxifen administration. Images are representative of the clones found in the 3 animals analysed per group. On the left panel, yellow highlights the contact area with the BM, i.e., the overlapping region between cells and BM reconstruction. Right panel: green highlights the clone in 3D. Scale bars: 5 μm. All statistical analyses were determined using the two-tailed Mann–Whitney test.
