Extended Data Fig. 6: RAP2 contributes to cell density-induced inactivation of YAP and TAZ.

a, Cytoplasmic translocation of YAP and TAZ caused by cell contact involves RAP2 and the Hippo pathway. Scale bars, 25 μm. MM-5KO, a HEK293A clone with deletion of MST1, MST2, MAP4K4, MAP4K6 and MAP4K7. RAP2-MST-KO, deletion of RAP2A, RAP2B, RAP2C, MST1 and MST2. The images are representative of three biologically independent experiments with similar results. b, Western blot showing absence of RAP2 proteins in RAP2-KO and RAP2-MST1/2-KO cells. Note that part of these results is shown in Fig. 1b and are from the same experiment. The image is representative of two independent experiments with similar results. c, Phosphorylation of YAP and TAZ induced by cell contact requires RAP2, MST1 and MST2. The western blot shows phosphorylation of YAP in cells with low, medium, or high confluence. The image is representative of two independent experiments with similar results. d, Deletions of RAP2A, RAP2B, RAP2C, MST1 and MST2 interfere with regulation of YAP and TAZ by cell density. Subcellular fractionations were performed for wild-type, RAP2-KO, MST1/2-dKO and RAP2-MST1/2-KO HEK293A cells cultured at low (S, sparse) or high density (D, dense). GAPDH and LaminB1 are markers for the cytoplasmic and nuclear fraction, respectively. The image is representative of two independent experiments with similar results. e, RAP2A, RAP2B, RAP2C, MST1 and MST2 are required for regulation of YAP and TAZ target genes by cell density. qPCR was performed to determine the expression of the YAP and TAZ target genes CTGF and CYR61 in the above cells at low or high confluence. Data are represented as mean ± s.e.m. n = 3 biologically independent samples.