Extended Data Fig. 5: Forced nuclear expression of YAP or TAZ induces canonical YAP/TAZ signaling in ECs.
From: A YAP/TAZ-TEAD signalling module links endothelial nutrient acquisition to angiogenic growth
a, Immunoblots of HUVECs transduced with doxycycline (Dox)-inducible control (Ctrl), YAPS127A (iYAPS127A) and TAZS89A (iTAZS89A) encoding lentiviruses, showing expression of FLAG-tagged YAPS127A and TAZS89A upon Dox treatment. Samples were analyzed 48 h after treatment with Dox or vehicle. b, RT-qPCR analysis of the canonical YAP/TAZ target genes ANKRD1, CTGF, and CYR61 in iYAPS127A and iTAZS89A expressing HUVECs. Expression changes are shown relative to Ctrl (n = 8 independent samples). c, d, Gene set enrichment analysis (GSEA) showing an enrichment of the YAP-conserved target gene expression signature in the transcriptomes of iYAPS127A (c) and iTAZS89A (d) expressing HUVECs. ES, enrichment score; NES, normalized enrichment score. e, f, FLAG immunoprecipitation studies in HUVECs overexpressing FLAG-tagged iYAPS127A and iYAPS94A/S127A (e) or iTAZS89A and iTAZS51A/S89A (f). The mutation of serine 94 to alanine in YAP and serine 51 to alanine in TAZ disrupts the interaction of (nuclear) YAP and TAZ with TEADs. g, Heatmap of mRNA expression changes in HUVECs expressing Ctrl, iYAPS127A, iYAPS94A/S127A, iTAZS89A or iTAZS51A/S89A as determined by RNA-seq. Canonical YAP/TAZ target genes are shown (n = 3 independent samples). Western blot data in a, e and f are from the respective experiment, processed in parallel, and are representative of at least three independent experiments. For b, data represent mean ± s.e.m.; two-tailed unpaired t-test. For c, d, Kolmogorov-Smirnov test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. The numerical data, unprocessed western blots and P values are provided as source data.
