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 uq^1ks (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 uq^1ks. C. Whole genome sequencing based mapping of mutant uq^2ks 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 uq^2ks. E. Lateral views of sibling and tmem2^hu5935 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 tmem2^hu5935 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 tmem2^hu5935 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 tmem2^hu5935 mutant embryos showing no difference in expression patterns of levels. Reciprocally, tmem2 expression was unchanged in vegfaa and kdr/kdrl morphants.