Fig. 5

gmfg mediates blood flow-dependent HSPC development. a Quantitative RT-PCR analysis of gmfg expression in FACS-sorted kdrl + cmyb + HE of control and tnnt2a-MO embryos at 48 hpf. Bars represent mean ± SD (n = 3). *p < 0.05. b Western blotting showing the protein level of Gmfg in the dissected trunk and tail of control and tnnt2a-MO embryos at 48 hpf. Representative blot is shown in the figure (Full-length blots are presented in Additional file 2: Fig. S3). Data represent mean ± SEM intensity of indicated blots (n = 3). *p < 0.05. c Representative images showing runx1/cmyb expression (red arrowheads) in the DA in control, tnnt2a-MO, and coinjection of tnnt2a-MO and gmfg mRNA embryos at 36 hpf. d Enumeration of runx1 + /cmyb + HSPCs shown in (c). Bars represent mean ± SD of control (n = 32), tnnt2a-MO (n = 43), and tnnt2a-MO + gmfg-mRNA (n = 23) embryos. ****p < 0.0001. e Representative images showing cmyb expression (red arrowheads) in the DA of control, tnnt2a-MO, and coinjection of tnnt2a-MO and gmfg mRNA embryos at 48 hpf. f Enumeration of cmyb + HSPCs shown in (e). Bars represent mean ± SD of control (n = 5), tnnt2a-MO (n = 10), and tnnt2a-MO + gmfg-mRNA (n = 10) embryos. ***p < 0.001, ****p < 0.0001. g Maximum projections of 80 hpf cd41:eGFP transgenic embryos injected with control MO, tnnt2a-MO, and tnnt2a-MO + gmfg-mRNA. Arrowheads denote cd41 + cells in the CHT. All views: anterior to left. h Enumeration of cd41 + cells shown in (g). Bars represent mean ± SD of control (n = 11), tnnt2a-MO (n = 12), and tnnt2a-MO + gmfg-mRNA (n = 11) embryos. **p < 0.01, ***p < 0.001. i FACS analysis showing the percentage of cd41 + cells in the dissected trunk and tail of control, tnnt2a-MO, and tnnt2a-MO + gmfg-mRNA embryos at 80 hpf (n = 3). ***p < 0.001, ****p < 0.0001. All scale bars, 100 µm