Fig. 1

vegfc loss-of-function results in decreased HSPC emergence from the DA. (A) Schematic showing the wave of emergence from the DA; this experiment occurs during the peak. (B) Single frame of time-lapse (hpf:min) showing cd41:gfp+ (green) HSPCs (arrows) budding from flk:mCherry+ (magenta) HE in Cas9 control (top panel) or vegfc crispant embryos (bottom panel). Dotted line outlines developing pronephric tubules (see Movies 1 and 2). (C) Quantification of the total number of cd41:gfp+ HSPCs budding from the DA between 48 and 52 hpf, divided by four to give cell number (#) per hour. n=5 embryos per condition, P=0.0019 (unpaired two-tailed t-test). (D) Four frames selected from Cas9 control and vegfc crispant time-lapse movies depicting HSPC budding (top panels) and a static hammock-like cell (bottom panels). White arrowheads indicate the cell of interest. Error bars show mean±s.e.m. Scale bars: 50 µm (B); 10 µm (D).