Fig. 3: YAP/TAZ-TEAD fuel endothelial mTORC1 activity by orchestrating the transcription of nutrient transporters.
From: A YAP/TAZ-TEAD signalling module links endothelial nutrient acquisition to angiogenic growth
a,b, Venn diagrams of up- (a) or downregulated (b) genes (log2 fold change (FC) ≥1 and FDR ≤ 0.05) in HUVECs transduced with inducible YAPS127A (iYAPS127A), TAZS89 (iTAZS89A) or control (Ctrl) lentiviruses as assessed by RNA-seq. c,d, Gene set enrichment analysis plots depicting an enrichment of genes associated with activated mTORC1 signalling in HUVECs expressing iYAPS127A (c) or iTAZS89A (d). ES, enrichment score; NES, normalized enrichment score. e, Heatmap of the enriched ‘mTORC1 signalling’ genes showing induction of these transcripts by iYAPS127A and iTAZS89A but not by the TEAD-binding-deficient iYAPS94A/S127A iTAZS51A/S89A mutants (n = 3 independent samples). f, Immunoblot analysis of S6K, S6 and 4EBP1 in Ctrl, iYAPS127A and iTAZS89A transduced HUVECs, assessing phosphorylation at mTORC1-sensitive sites. g,h, Phosphorylation status of S6K, S6 and 4EBP1 in HUVECs that were transfected with siRNAs targeting YAP/TAZ (siYAP/TAZ) (g) or TEAD1/TEAD2/TEAD4 (siTEAD1/2/4) (h). i,j, Immunolabelling of p-S6Ser235/236, VECAD and PECAM in P6 retinas of Ctrl, TaziEC-GOF (i) and Yap/TaziEC-KO (j) mutants. Scale bars, 200 μm. The isolated p-S6Ser235/236 signal is shown in grey at the bottom. Arrows indicate the peri-venous region in TaziEC-GOF (white) and Yap/TaziEC-KO mice (transparent). k, Heatmap of solute carrier expression in Ctrl, iYAPS127A-, iYAPS94A/S127A-, iTAZS89- or iTAZS51A/S89A-expressing HUVECs determined by RNA-seq (n = 3 independent samples). l,m, Immunoblot analysis of SLC7A5 and SLC1A5 in siYAP/TAZ (l) or siTEAD1/2/4 (m) transfected HUVECs. n, SLC7A5 and SLC1A5 protein levels in HUVECs expressing Ctrl, iYAPS127A, iTAZS89A, iYAPS94A/S127A or iTAZS51A/S89A. o, TEAD-depleted HUVECs fail to induce SLC7A5 and SLC1A5 in response to iYAPS127A or ITAZS89A overexpression as determined by immunoblotting. p, Analysis of endothelial YAP, TAZ and TEAD1 ChIP–seq peaks revealed the TEAD-binding sequence as a highly enriched motif. q,r, TAZ, YAP and TEAD1 ChIP–seq signals at the SLC7A5 (q) and SLC3A2 (r) genomic loci. RPKMs, reads per kilobase per million mapped reads. Western blot data in f–h and l–o are from the respective experiment, processed in parallel and are representative of at least three independent experiments. For c and d, the Kolmogorov–Smirnov test was used. The unprocessed blots are provided as source data.
