Skip to main content
. Author manuscript; available in PMC: 2016 May 7.
Published in final edited form as: Cell. 2015 May 7;161(4):858–867. doi: 10.1016/j.cell.2015.04.017

Figure 6.

Figure 6

mtClpX is important for vertebrate heme biosynthesis and erythropoiesis. (A) Relative mRNA abundance for human CLPX, CLPP, ALAS2, and SLC25A38 (indicated as S25A38) throughout erythropoiesis as indicated in a microarray dataset described in (Novershtern et al., 2011). Erythroid development stages were defined by cell type specific markers: 1, CD34+ CD71+ GlyA−; 2, CD34− CD71+ GlyA−; 3, CD34− CD71+ GlyA+; 4, CD34− CD71low GlyA+; 5, CD34− CD71− GlyA+. See also Fig. S5A,B for expression in MEL cells and zebrafish embryos. (B) o-dianisidine staining (brown) for hemoglobinized red cells in zebrafish embyos. Embyros were grown from zygotes injected at the 1-2 cell stage with clpxa-targeting morpholinos or uninjected zygotes (control). (C) Erythrocyte development at 72 h post-fertiliztion (hpf) was quantified by flow cytometry, using dissociated cells from Tg(globin-LCR:eGFP) zebrafish. P ≤ 0.01 for erythrocyte reduction by clpxa knockdown with either morpholino. See also Fig. S5C-E for qPCR quantitation of clpxa knockdown and nontargeting morpholino injections. (D) Rescue of clpxa MOb-induced anemia by ALA supplementation. Tg(globin-LCR:eGFP) zebrafish embryos were supplemented with 2 mM ALA from 24 to 72 hpf, upon which GFP+ erythrocytes were quantified by flow cytometry. p = 0.025 for rescue of anemia in clpxa knockdown embryos by ALA supplementation (E) Heterozygous sauternes or frascati zebrafish were crossed, and progeny were grown for 72 hpf, with or without ALA supplementation as in (D). Anemia was assayed by o-dianisidine staining. p = 0.04 for rescue of anemia in sauternes+/− progeny by ALA. n=52 for sauternes −ALA; n=43 for sauternes +ALA; n=98 for frascati −ALA; n=122 for frascati +ALA. Error bars represent mean ± SD.