Figure 3.

Loss-of-Function Analysis of the RBPMS, RBPMS2, and GTF2E2 Orthologs in Zebrafish

After injection of 0–3 ng ATG and splicing morpholinos (MOs) against the RBPMS zebrafish ortholog (row E), both the o-dianisidine/benzidine staining (arrows) in embryos at 48 hpf (right) and the embryonic βe3 globin expression in embryos at 16–18 ss (left) are obviously decreased, indicating a dose-dependent disruption in erythropoiesis in the experimentally treated embryos as compared to uninjected and gtf2e2-, rbpms2a-, and rbpms2b-MO-injected controls (rows A–D). Representative results are shown for the embryos injected with MOs against the RBPMS ortholog in (E) as well as for the embryos injected with MOs against rbpms2a (C) and rbpms2b (D) at higher doses. Injections of MO against the zebrafish GTF2E2 ortholog (B) also at a higher dose show no obvious effect on βe3 globin expression at 16–18 ss and o-dianisidine/benzidine staining at 48 hpf. Expression pattern of vascular marker gene kdrl (A–E, middle) is relatively normal in all MO-injected embryos at 24–26 hpf, suggesting grossly normal development of cells in other organs. The numbers on the lower right corner of each image indicate the number of embryos with phenotypes similar to the ones shown on each of the images over the total number of embryos examined in each of the experimental groups.