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Fig. S1
Related to Figure 1: The Vegf pathway and tmem2 do not regulate one another at the transcriptional level.
A. Whole genome sequencing based mapping of mutant uq1ks (derived from a forward genetic screen for venous sprouting defects) showing linkage to a region of Chromosome 5 containing tmem2. Sequence analysis identified a truncating mutation (Q616X) in tmem2. B. Protein schematic showing the predicted resultant protein in uq1ks. C. Whole genome sequencing based mapping of mutant uq2ks showing linkage to a region of Chromosome 5 containing tmem2. Sequence analysis identified a mutation in intron 6 (IVS6-9T>A) affecting the splice acceptor site in exon 7. D. Protein schematic showing the predicted resultant protein in uq2ks. E. Lateral views of sibling and tmem2hu5935 mutant trunks stained by ISH for arterial (hey2; n=38) and venous (dab2; n=46) markers showing no alteration in expression. F. Lateral views of sibling and tmem2hu5935 mutants on the Tg(kdrl:eGFP) background showing mild (>15 ISVs), medium (5- 15 ISVs) and severe (<5 ISVs) phenotypes. Scale bar = 100 μm G. Pie charts showing significantly greater severity of phenotype in tmem2hu5935 mutants following suboptimal Vegf inhibitor (SU5416) treatment compared with DMSO treatment (p<0.001 by Chi-square analysis; n=37 DMSO, n=46 SU5417), according to the categories depicted in F. H. ISH expression analysis for Vegf pathway members vegfaa, kdrl and flt4 in sibling and tmem2hu5935 mutant embryos showing no difference in expression patterns of levels. Reciprocally, tmem2 expression was unchanged in vegfaa and kdr/kdrl morphants.
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