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. Author manuscript; available in PMC: 2022 Sep 7.
Published in final edited form as: Circulation. 2021 Jun 29;144(10):805–822. doi: 10.1161/CIRCULATIONAHA.120.053047

Figure 7. YAP/TAZ inhibition improves AVM formation in Alk1 mutant retinas.

Figure 7.

(A) YAP and TAZ staining of siCon, ALK1, SMAD4 or ENG siRNAs transfected HUVECs treated with DMSO or Verteporfin (VP, 5 μM) for 6 h. Nuclear YAP/TAZ localization in siALK1, siSMAD4 or siENG ECs is blocked by VP treatment. A scale bar: 50 μm. (B) Quantification of nuclear YAP and TAZ from siCon, siALK1,siSMAD4 and siENG transfected HUVECs. ***P<0.001, ns: nonsignificant, Two-way ANOVA with Tukey’s multiple comparisons test. (C) Experimental strategy to assess the effects of YAP/TAZ inhibition in EC specific Alk1 deleted vasculature. Arrowheads indicate the time course of Tx (100 μg) and VP (50mg/kg) or vehicle administration. (D) IB4 staining of P6 retinal flat mounts from VP injected Alk1f/f, Alk1f/f CDH5 CreERT2 or Alk1f/f Mfsd2a CreERT2 mice. (E) Stereomicroscopy images of vehicle or VP injected Alk1f/f Mfsd2a CreERT2 retinas. (F) Quantification of the AVM number/retina. Each dot represents one retina. n = 6–8 retinas per group. Error bars: SEM. ***P-value < 0.001, One-way ANOVA with Sidak’s multiple comparisons test. (G) Angular histograms showing polarization angles of artery and vein from Alk1f/f Mfsd2a CreERT2 with VP. (H) PI box plots of Alk1f/f Mfsd2a CreERT2 with vehicle or VP. n=5–6 retinas, Error bars: SEM, ***P-value < 0.001, ns: nonsignificant, Two-way ANOVA with Tukey’s multiple comparisons test. Scale bars : 50 μm (A), 500 μm (D), 300 μm (E)