ZFIN Figure: Savage et al., 2019, Fig. 7

Fig. 7

SOCE inhibition limits sprouting angiogenesis and disrupts signalling pathways downstream of VEGF. a, b SKF9635 impairs angiogenesis in Tg(fli1a:EGFP) embryos (white arrowheads) and induces abnormal tip cell morphology (blue arrowhead) by 27 hpf. Scale bars 50 µm. c SKF-96365-treated embryos display reduced SeA length by 27 hpf (unpaired t-test; p = < 0.0001; t = 11.22 DF = 14; 2 repeats, n = 4 embryos per group). d, e SKF-96365 treatment impairs DLAV formation (white asterisks) in comparison with control embryos, which form the DLAV normally by 50 hpf (white arrowheads). Abnormal tip cell morphology (blue arrowhead) remain in SKF-96365-treated embryos by 50 hpf. Scale bars 50 µm. f SKF-96365-treated embryos display increased frequency of abnormal vascular branching (stunted SeAs or incomplete DLAV anastomosis) by 50 hpf (unpaired t-test; p = < 0.0001; t = 14.28 DF = 18. n = 2 repeats, n = 5 embryos per group). g tmem33 expression is not significantly altered by SKF-96365 treatment (unpaired t-test; t = 0.911, DF = 4, n = 3 repeats). h–j SKF-96365 reduces frequency of endothelial Ca²⁺ oscillations in Tg(fli1a:gal4FF^ubs3;uas-gcamp7a) embryos. Intensity projection over time showing calcium oscillations in control embryos, highlighting intensity change over duration of time lapse (unpaired t-test; p = < 0.05; t = 2.594, DF = 10, n = 2 repeats, 3 embryos per group). k, l Notch reporter expression is reduced in SeAs (white arrowheads) and DA (red arrowhead) of Tg(csl:venus) embryos. Scale bars 50 µm. m DA expression of Notch reporter is reduced by SKF treatment (unpaired t-test, **p = 0.0097, t = 3.982 DF = 15, 2 repeats, n = 4 embryos per group). n, o SKF-96365 treatment reduces endothelial filopodia number (black arrowheads) and abnormal spade-shaped tip cell morphology (blue arrowhead), in Tg(fli1a:lifeact-mClover) embryos. Scale bars 5 µm. p SKF-96365 treatment reduces mean filopodia number per SeA per embryo by 26 hpf (unpaired t-test; p = < 0.05; t = 2.525, DF = 10, n = 2 repeats, n = 5 embryos per group). Source data are provided as a Source Data file

Expression Data

Genes:	EGFP GCaMP mClover tmem33 Venus
Fish:	WT qmc61Tg sh467Tg y1Tg sh392Tg; ubs3Tg
Condition:	chemical treatment by environment: SKF-96365 hydrochloride
Anatomical Terms:	dorsal aorta blood vessel endothelial cell dorsal longitudinal anastomotic vessel blood vessel endothelial cell endothelial tip cell filamentous actin intersegmental vessel blood vessel endothelial cell intersegmental vessel endothelial tip cell whole organism
Stage Range:	Prim-5 to Long-pec

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	qmc61Tg sh467Tg y1Tg sh392Tg; ubs3Tg
Condition:	chemical treatment by environment: SKF-96365 hydrochloride
Observed In:	dorsal aorta blood vessel endothelial cell dorsal aorta Notch signaling pathway dorsal longitudinal anastomotic vessel dorsal longitudinal anastomotic vessel sprouting angiogenesis endothelial tip cell calcium-mediated signaling endothelial tip cell filopodium endothelial tip cell regulation of cytosolic calcium ion concentration endothelial tip cell regulation of store-operated calcium channel activity intersegmental vessel intersegmental vessel blood vessel endothelial cell intersegmental vessel endothelial tip cell intersegmental vessel endothelial tip cell filopodium assembly intersegmental vessel sprouting angiogenesis
Stage Range:	Prim-5 to Long-pec

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.