Fig 2: Maturation of leaflets and commissures. (A) Immunolocalization at P6 of Nfatc1 (green) and Prox1 (red) in VV of WT mice administered control solvent or cyclosporin (n = 5 versus 8 VVs, P < 0.05, ?2). (B) Immunolocalization of Foxc2 (green) and Prox1 (red) in VVs of mice of the indicated genotypes. n = 6 per condition. (C and D) Quantification of leaflet length and stage of VV development in mice of the indicated genotypes. Error bars represent means ± SEM. t test for leaflet length. (C, D, G, H, and I) Colors represent developmental stage (see key). The number of VVs analyzed for each condition is given above each bar. *, P < 0.05; **, P < 0.005 (?2 for developmental stage). (E) Immunolocalization of Cx37 (arrowheads) and Cx43 in VV leaflets at P6. n = 4. (F) Localization of Prox1, Foxc2, and SMA at P6 in Gja4-/- vein. n = 6 (G and H) Stages of VV development reached in mice of the indicated genotypes. (I) Localization of Prox1, GFP, and SMA in Gjc2GFP/GFP KO reporter vein at P2 (n = 4), and stages of VV development reached in mice of the indicated genotypes. (J) Localization of Cx47-expressing VV cells (arrowheads) of Gjc2GFP/+ reporter mice at P6. n = 4. Multichannel images are reproduced in supplementary data. Bars, 20 µm. (A–J) Blue stain is PECAM1.