Fig. 2

Increased, Premature, and Prolonged LEC Specification in Nova2 Mutants

(A and B) Immunofluorescence (IF) staining of Prox1 at 28, 30, 32, 36, and 54 hpf in Tg(fli1a:nEGFP)^y7 siblings and nova2^uq14bh mutants. Arrowheads indicate Prox1-positive ECs (brackets indicate PCV). Left hand images show merged zoom out of trunk as an overlay of Prox1 and fli1a:nEGFP, which was used to confirm nuclear localization.

(C) Quantification of the number of Prox1-positive ECs at 28 hpf (siblings n = 14 and mutants n = 11), 30 hpf (siblings n = 23 and mutants n = 22), 32 hpf (siblings n = 18 and mutants n = 15), 36 hpf (siblings n = 40 and mutants n = 17), and 54 hpf (siblings n = 15 and mutants n = 15). For all stages, 7 body segments were quantified except at 54 hpf when 6 body segments were used (see also Figures S3A–S3E).

(D) Quantification of the number of Prox1-positive ECs in the PCV wall or having sprouted dorsally in 30 hpf sibling and mutant embryos. n = 7 Prox1-positive ECs in 4/23 embryos for siblings, and n = 33 Prox1-positive ECs in 14/22 embryos for mutants, with 11/33 Prox1-positive ECs sprouting from the PCV.

Error bars represent SD, Mann-Whitney-test (C), ^∗p < 0.05 ^∗∗p < 0.01, and ^∗∗∗p < 0.001; Scale bar represents 25 μm in (A) and (B).

See also Figure S3.