Extended Data Fig. 3: CCM3 regulates YAP/TAZ activation independently of Hippo signalling.
a, Immunoblotting analysis of YAP, TAZ, pLATS1/2, pS109-YAP, and pS127-YAP in human CAFs. The scatter plots show quantification of YAP, TAZ, pLATS1/2, pS109-YAP, and pS127-YAP relative to siRNA control-transfected human CAFs. Each data point represents an independent experiment. n: independent experiments (YAP: HN-CAF, 6; V-CAF, 10; Cer-CAF, 6. TAZ: HN-CAF, 4; V-CAF, 4; Cer-CAF, 3. pS127-YAP: HN-CAF, 6; V-CAF, 10; Cer-CAF, 6. pS109-YAP: HN-CAF, 5; V-CAF, 10; Cer-CAF, 5. pLATS1/2: HN-CAF, 6; V-CAF, 10; Cer-CAF, 6). Line and error bars indicate mean ± s.d.; one-way ANOVA test. b, Immunofluorescence analyses of YAP/TAZ in V-CAFs transfected with siRNA targeting various components of the STRIPAK complex and stained for YAP/TAZ (green), F-actin (red), and DAPI (blue). The scatter plots show quantification of the YAP/TAZ nuclear/cytoplasmic ratio. Each data point represents individual cells from three independent experiments. n: cells (siCtr, 44; siSTRIP1, 48; siSTRIP2, 48; siSLMAP, 26; siCCM3, 46; siMST3, 47; siMST4, 46; siSOK1, 28; siPPP1R14C, 22). Line and error bars indicate mean ± s.d.; one-way ANOVA test. Scale bars, 50 μm. c, Immunoblotting analysis of pS127-YAP and pLATS1/2 in V-CAFs after serum starvation for 30 and 60 min. d, Immunofluorescence analysis of YAP/TAZ localization after serum starvation for 30 and 60 min in V-CAFs. e, Immunofluorescence analysis of YAP/TAZ localization in dense cultures of V-CAFs. f, Immunofluorescence analysis of YAP/TAZ localization in V-CAFs, HN-CAFs and Cer-CAFs plated on different concentrations of collagen-gels. Quantification of the nuclear/cytoplasmic YAP/TAZ ratio in V-CAFs plated on different concentrations of collagen-gels. n > 17 individual cells. Line and error bars indicate mean ± s.d. Scale bars, 50 μm. Statistical data and uncropped blots are provided in the source data.
