Fig. 4

tmem33 functions downstream of Vegfa during angiogenesis. a–e tmem33 knockdown prevents Vegfa-mediated increases in Ca²⁺ oscillations. Overexpression of vegfa₁₆₅ significantly increases frequency of Ca²⁺ oscillations in SeAs, whereas tmem33knockdown alone and in the presence of vegfa₁₆₅ significantly reduce frequency of Ca²⁺ oscillations in SeAs. Intensity projection over time showing Ca²⁺ oscillations highlighting intensity change over duration of time lapse. Embryos display changes in fluorescence in SeA tip cells (white arrowheads) (two-way ANOVA with Holm–Sidak’s corrections: *p = < 0.05. 2 Repeats, n = 3 embryos per group) (scale bars 50 µm). f–h tmem33 knockdown using 0.4 ng morpholinos reduces EC number in ISVs at 53 hpf. h Unpaired t-test; ****p < 0.0001; t = 6.507; df = 30; 2 repeats; n = 8 embryos per group. i VEGF receptor expression is not significantly affected in tmem33 crispants (one-way ANOVA with Dunnett’s corrections; p = < 0.05, n = 3 repeats). Source data are provided as a Source Data file