Fig. S5

5-Aminolevulinic acid does not restore hemoglobin levels to alas2 or slc25a38a+b morphants.Zebrafish embryos at 48-hpf were stained for hemoglobin with o-dianisidine following microinjection (at 1-cell stage) with the alas2 or control standard MO. A) The typical phenotypes of control standard morpholino (STD MO) and alas2 morpholino-injected embryos stained with o-dianisidine are shown. B) The morphants were incubated with or without 0.3 mM of 5-Ala added to the egg water at 4 hours post-injection. The level of o-dianisidine hemoglobin staining was scored visually as “low”, “medium” or “normal” in a blinded manner and the proportions of embryos in each category are presented in the graph. Results of three separate experiments are presented with the actual numbers of scored embryos indicated on the right hand axis of the graph. There was no statistically significant difference between untreated and 5-Ala-treated morphants indicating that 5-Ala supplementation did not increase hemoglobin level.