Extended Data Fig. 2: Expression and subcellular localization of TAZ in angiogenic ECs.
From: A YAP/TAZ-TEAD signalling module links endothelial nutrient acquisition to angiogenic growth
a, RNA-seq analysis of YAP and TAZ transcript levels in different human EC cultures (n = 3 independent samples). HAECs, human aortic ECs; HMVECs, human microvascular ECs; HUVECs, human umbilical vein ECs; HDLEC, human dermal lymphatic ECs. b, Immunoblot analysis of YAP and TAZ protein expression in different endothelial subtypes. c, Targeting strategy to generate a Taz (Wwtr1) knock-in allele encoding for a GFP, FLAG, and biotin-labelling peptide tagged TAZ protein. A schematic representation of the wild-type Taz (Wwtr1) locus, the targeting vector, the recombined as well as the excised locus is shown. E5−7, exons 5 to 7. Triangles denote loxP sites. Neo, neomycin positive selection cassette. GFB, GFP-FLAG-biotin labelling peptide fusion tag. DTA, diphteria toxin negative selection marker. d, PCR analysis of genomic DNA obtained from wild-type, heterozygote and homozygote Taztag mice. Lane 1, DNA marker. e, RT-qPCR analysis of the canonical YAP/TAZ target gene Ctgf in retinal ECs derived from wild-type (Ctrl) and homozygous Taztag/tag mice, showing no expression difference between the two genotypes (n = 2 (Ctrl) and 3 (Taztag/tag) independent samples). f, g, ERG- and PECAM- immunofluorescence staining (f) and quantification of vascular parameters (g) of P6 Ctrl and Taztag/tag retinas, revealing no gross difference in vascular morphogenesis between the two genotypes (EC area: n = 4 (Ctrl) and 8 (Taztag/tag) independent samples; Number of ECs: n = 3 (Ctrl) and 9 (Taztag/tag) independent samples). h, Images of the vascular front and plexus in TAZ, ERG and PECAM labelled P6 mouse retinas derived from C57BL/6 wild-type mice. The images in grey (right panels) show the isolated TAZ signal. Western blot data in b are from the respective experiment, processed in parallel, and are representative of at least three independent experiments. For a, e and g, data represent mean ± s.e.m.; two-tailed unpaired t-test. **P < 0.01; ****P < 0.0001; NS, not significant. The numerical data, unprocessed western blots and P values are provided as source data.
