Fig. S3

Trpp2 and Trpv4 are essential for valve development. (A-B) Tg(klf2a:H2B-eGFP) relative expression level (A) and relative number of cells counted in the AVC (B) in controls, trpp2, trpv4 and trpv4+trpp2 MO embryos (n=6, 7, 7 and 4, respectively). (C) Relative number of cells counted in the AVC in controls, trpp2^cup-/-, and trpv4^sa1761-/- at 48 hpf based on Tg(fli1:nls-mcherry) nuclei number in the AVC. (D) Total number of valve cells counted in the AVC in controls, trpp2^cup-/-, and trpv4^sa1761-/- at 80 and 96 hpf based on Tg(fli1:nls-mcherry) nuclei number in the AVC (see also MovieS4). (E) Single embryo follow up of the valve defects associated with early decrease of cells in AVC. For the trpv4 knock down embryos, 42% (n=5) displayed a 50% decrease of cells in the AVC. All of these embryos had upper and lower valve defects at 96 hpf as described in Figure 3H. For the trpp2 mutant embryos, 55% (n=5) displayed a 32% decrease of cells in the AVC. All of these embryos had upper and lower valve defects at 96 hpf as described in Figure 3H. (F) Table summarizing the valve rescue obtained after overexpression of trpp2 and trpv4 specifically in the endocardium of gata2 knock down embryos. (F) Table summarizing the lack of valve rescue obtained after gata1 knock down in trpp2^cup-/- and trpv4 knock down at 96 hpf. (H) Typical phenotypes observed at 80 hpf and 96 hpf to score the rescue of the valve phenotype in gata1 and gata2 knock down. We analyzed Tg(fli:kaede) embryos incubated with bodipy-fl to label the leaflets.