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Fig. 7
Restoration of ISV Sprouting by HA'se Requires Vegf Signaling, and Vegf Signaling Is Sufficient to Rescue the tmem2 Mutant Phenotype
(A) Experimental strategy for HA'se and Vegf pathway epistasis experiments.
(B) Scatterplots demonstrating that, as observed in Figure 6D, the significant reduction in ISV number in tmem2 mutant embryos (n = 14) compared with siblings (n = 11) is rescued when HA'se is injected into mutants (n = 16) compared with siblings in the presence of DMSO (n = 18). When embryos are incubated with SU5416 from the time of injection, tmem2 mutants (n = 21) have significantly fewer ISVs than siblings (n = 10) when injected with dextran but tmem2 mutants (n = 21) also have significantly fewer ISVs than siblings (n = 9) when injected with HA'se, demonstrating that rescue by HA'se requires intact Vegf signaling.
(C) Lateral view images of sibling and tmem2hu5935 mutants on the Tg(kdrl:EGFP) background either negative for Tg(hsp70l:gal4)/Tg(10xuas:vegfc) (control) or positive for Tg(hsp70:gal4)/Tg(10xUAS:vegfc) (vegfc O/E). Scale bar, 100 μm.
(D) Scatterplot showing number of ISVs in sibling (open circles; n = 10) and tmem2hu5935 mutants (red squares; n = 22) negative for vegfc O/E, showing a significant reduction in ISV number. tmem2hu5935 mutants positive for vegfc O/E (green triangles, n = 10) had significantly more ISVs than tmem2hu5935 mutants negative for vegfc O/E and were not significantly different from siblings negative for vegfc O/E.
∗∗p < 0.01, ∗∗∗∗p < 0.0001; ns, not significant. Error bars represent ±SD.
Reprinted from Developmental Cell, 40, De Angelis, J.E., Lagendijk, A.K., Chen, H., Tromp, A., Bower, N.I., Tunny, K.A., Brooks, A.J., Bakkers, J., Francois, M., Yap, A.S., Simons, C., Wicking, C., Hogan, B.M., Smith, K.A., Tmem2 Regulates Embryonic Vegf Signaling by Controlling Hyaluronic Acid Turnover, 123-136, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell