Extended Data Fig. 8: Exogenous CCM3 regulates differentiation cell fate of MSCs.
a, Immunofluorescence analysis of the basal plasma membrane of MSCs stably expressing GFP, CCM3–GFP, CCM3-mutant-GFP or FAT–GFP. Scale bar, 20 μm. b, Immunoblotting analyses against αSMA, pS19-MLC2 and tubulin. c, MSCs expressing exogenous GFP-tagged constructs were stained for YAP/TAZ and the nuclear/cytoplasmic ratio quantified. Each data point represents an individual cells. n = 20 cells. Line and error bars indicate mean ± s.d.; one-way ANOVA test, following up multiple comparisons. Scale bars, 20 μm. d, MSCs stably expressing exogenous CCM3 constructs were plated on soft (1 kPa) and stiff (50 kPa) collagen-coated softwells. The MSCs were differentiated into osteocytes. The calcium deposition was visualized with Alizarin red staining. Each data point represents individual images quantified from three independent experiments. n = 6 individual images. Line and error bars indicate mean ± s.d.; one-way ANOVA test. e, MSCs stably expressing exogenous CCM3 constructs were plated on soft (1 kPa) and stiff (50 kPa) collagen-coated softwells. The MSCs were differentiated into adipocytes. Adipogenic differentiation were stained with oil red O to detect lipids. Each data point represents individual images quantified from three independent experiments. n = 6 individual images. Line and error bars indicate mean ± s.d.; one-way ANOVA test. Statistical data and uncropped blots are provided in the source data.
