Extended Data Fig. 9: Characterizing the effect of phc1 overexpression on Notch signalling.
a-l, WISH of WT sibling and supt16h-/- embryos injected with phc1-MO probed for notch1a, dla, and dlc at 28 hpf. m-o, Representative confocal images of wild-type embryos on a Tg(Tp1:GFP) background injected with 100 and 200 pg of phc1 mRNA at 28 hpf. Insets are magnified images of the DA and intersomitic vessels. Representative images from one independent experiment. p-c’, WISH of wild-type embryos injected with 100 pg of phc1 mRNA probed for at 28 hp for runx1 (p-q), phc1 (r-s), notch1b (t-u), notch3 (v-w), dla (x-y), dlc (z-a’), and dll4 (b-c’). d’, RT-qPCR of Notch genes in WT sibling and embryos injected with 100 pg (r) of phc1 mRNA collected at 28 hpf. No significant changes observed between samples. (Represented as mean ± s.d., two-tailed Student’s t-test with Holm-Sidak correction for multiple comparisons, n = 3). e'-j’, WISH at 28 hpf for phc1 (e’-f’), runx1 (g’-h’), and notch1b (i’-j’) of wild-type embryos injected with fli1ep:phc1 plasmid for transient expression of phc1 in the vasculature. Blue arrowheads indicate HSPCs. Bar, 100 μm.
