Fig. 6: CAFs compression restrains cancer cell growth.

a Evolution of cancer cell clusters that freely grow (left) or grow compressed by CAFs (right). Cell nuclei are labeled with Hoestch to allow live imaging. White line outlines the boundary of the cancer cell cluster. Graphs: Quantification of the evolution of cancer cell number (left) and density (right). n = 6 clusters per condition from N = 2 independent experiments. b Immunostaining of cells in M-phase (labeled by Phospho-histone H3) in cancer cell clusters that are freely growing, confined or surrounded by control or MyosinIIA KO CAFs. Graph: quantification of the percentage of PHH3⁺ cells in each condition. Data are presented as mean ± SD, n = 18 (free growth), 15 (confined), 17 (CAFs control), and 17 (CAFs Myosin IIA KO), from N = 3 independent experiments. *p < 0.05, **p < 0.01, Kruskal–Wallis ANOVA test, Dunns multiple comparisons test. c Quantification of percentage of PHH3⁺ cells in control and αSMA:MyosinIIA^-/- tumors. Data are presented as mean ± SD, n = 6 (Control), 4 (αSMA:MyosinIIA^−/−), from N = 1 experiment. Not significative, Mann Whitney two-tailed test. d Immunostaining of apoptotic cells (labeled by cleaved-caspase 3) in cancer cell clusters that are freely growing, confined or surrounded by control or MyosinIIA KO CAFs. Graph: quantification of CC3+ area normalized to the total number of cancer cells per cluster. Data are presented as mean ± SD, n = 9 (free growth), 6 (confined), 11 (CAFs control), and 8 (CAFs Myosin IIA KO), from N = 2 independent experiments. e Quantification CC3+ area in control and αSMA:MyosinIIA^−/− tumors. Data are presented as mean ± SD, n = 5 (Control), 4 (αSMA:MyosinIIA^−/−), from N = 1 experiment. Representative examples of immunostainings quantified in c and e can be found in Supplementary Data Fig. 15. Scale bars = 100 µm.