Loss of Wdr26 blocks terminal erythropoiesis. (A) Western analysis confirmed the silencing of Wdr26 by shRNAs in primary mouse erythroblasts. Knockdown of Wdr26 led to reduced (B) enucleation, (C) hemoglobin production, and (D) expression of erythroid-induced genes in mouse primary erythroid progenitors. Error bars represent SEM from 3 replicates. *P < .05, **P < .01. (E) Silencing of Wdr26 led to increased R3 subpopulation and reduced R4 and R5 subpopulations in primary mouse erythroblasts. Error bars represent SEM from 3 replicates. *P < .05, **P < .01. (F) Western analysis confirmed the knockout of Wdr26 in MEL cells. (G) Porphyrin fluorescence assay and (H) o-dianisidine staining showed reduced heme production in differentiating MEL cells. Error bars represent SEM from 3 replicates. *P < .05, **P < .01. (I) Strategy to knock out wdr26b in zebrafish using 2 gRNA targeting sites (arrowheads) and verification by PCR using primers indicated as arrows. (J) Red blood cell number and (K) heme content in the peripheral blood of wdr26b^−/− fish were reduced in comparison to the wild-type fish. Error bars represent SEM from 6 animals. **P < .01. (L) The wdr26b-knockout fish showed reduced heme content in the adult hematopoietic tissue kidney. Error bars represent SEM from 5 animals. **P < .01. (M) Survival curve of wild-type (n = 6) and wdr26b^−/− (n = 6) fish under hypoxic condition.