Fig. 4
Morpholino Knockdown of Candidate Genes in Zebrafish Results in Profound Anemia
WT zebrafish embryos were injected at the one-cell stage with the respective morpholinos and stained at 48 hpf with o-dianisidine to detect hemoglobinized cells.
(A) Uninjected (WT) embryos show normal hemoglobinization as indicated by dark brown staining on the yolk sac (arrow).
(B–G) Morpholino-injected embryos. Accurate morpholino gene targeting was verified by RT-PCR (slc25a39, slc22a4, slc22a5, tmem14c, and c1orf69) or real-time quantitative RT-PCR (isca1) on cDNA from uninjected (WT) or morpholino-injected (mo) embryos. β-actin (actb) was used as a control for off-target effects in the RT-PCR. For RT-PCR, (ctrl) indicates no cDNA template control. For real-time quantitative RT-PCR, (ctrl mo) indicates embryos injected with a standard control morpholino.
Reprinted from Cell Metabolism, 10(2), Nilsson, R., Schultz, I.J., Pierce, E.L., Soltis, K.A., Naranuntarat, A., Ward, D.M., Baughman, J.M., Paradkar, P.N., Kingsley, P.D., Culotta, V.C., Kaplan, J., Palis, J., Paw, B.H., and Mootha, V.K., Discovery of genes essential for heme biosynthesis through large-scale gene expression analysis, 119-130, Copyright (2009) with permission from Elsevier. Full text @ Cell Metab.