Fig. 4

Hematopoietic stem and progenitor cells (HSPCs) are reduced in ism1 knockdown embryos.

Single-cell-stage embryos were injected with 7 ng of ism1 MO (MO), or 7 ng of ism1 MO and 17.88 ng of ism1 mRNA (Rescue); uninjected embryos served as a control. (A) Ten 48 hpf embryos from each treatment were digested, plated in methylcellulose with exogenous hematopoietic-supportive growth factors, and incubated for 7 days. (B) Colony forming units (CFUs) generated by ism1 MO (light blue) and ism1 MO with ism1 mRNA (Rescue; darker blue) injected embryos presented as fold change over control embryos (dark blue). (C) qRT-PCR was performed comparing transcript levels of cmyb and runx1 in ism1 MO injected (MO) and uninjected controls (control) at 48 hpf. Bars represent the mean, and error bars represent standard deviation. A two-tailed unpaired Student’s T test was performed to determine statistical significance. * denotes p<0.05, ** denotes p<0.0001, N.S indicates no significance.