Fig. 4

POFUT1 regulates coronary artery formation via DLL4/NOTCH1 signaling. a, b Co-staining for N1ICD and IB4 showing reduced expression of N1ICD in coronary plexus of E17.5 Pofut1 ^cKO heart. Dotted line separates trabeculae from compact myocardium. c DLL4-Fc binding and NOTCH1 extracellular domain antibody binding to control (blue profiles) and Pofut1 knockout (Pofut1 KO) (red profiles) CHO cells showing reduced DLL4 binding by Pofut1 KO cells, which have normal NOTCH1 expression. Grey profiles reflect secondary antibody alone. d, e PECAM1 and VEGFR3 staining for coronary angiogenic cells (arrowhead) in coronary plexuses at E12.5, one-day after Dll4 deletion, showing increased PECAM1+ area, the number of PECAM1+ tip-like or VEGFR3+ cells in Dll4 ^cKO (Dll4f ^/f ;Cdh5 ^CreERT2). f PECAM1 and VEGFR3 staining for coronary angiogenic cells (arrowhead) and arteries (arrow) at E15.5, one-day after Dll4 deletion, shows increased number of VEGFR3^high cells in Dll4 ^cKO. *p < 0.001. g EdU labeling of VEGFR3+ coronary angiogenic cells at E15.5, 1 day after Dll4 deletion, shows increased proliferation of VEGFR3 cells (arrowheads). All bar charts represent mean ± SD; *p < 0.001. en/ep, endocardium/epicardium