Fig 1: Fetal hematopoiesis is regulated by a redox sensitive complex of translation re-initiation factors Denr and Mcts1.a Heatmap depicting average oxidation level (%) of cysteine peptides that belong to translation re-initiation factor Denr in fetal and adult HSPCs. Statistics are shown in Table 1. Denr protein sequence with higher oxidized cysteines in fetus (red) and adult (blue) is shown. b Confocal images of in situ PLA foci (top) of Denr and Mcts1 translation re-initiation factors and proximity ligation assay (bottom) of Denr-Mcts1 complex in fetal and adult HSPCs using anti-Denr and anti-Mcts1 antibodies, and 4',6-diamidino-2-phenylindole (DAPI) for DNA staining. Graph (right) depicts number of PLA foci per ten FL and ABM HSPCs. c Zinc release from zinc finger binding domain of Denr upon incubation with thiol reactive oxidative agents H2O2, nitrosylated glutathione (GSNO), and glutathione disulphide (GSSG). Each point represents the mean of three measurements. d Heatmap with log2 fold change expression level fetal/adult for targets of Denr-Mcts1 mediated translation re-initiation (Schleich et al., 2017) with significantly higher protein levels in fetal HSPCs compared to adult HSPCs (Jassinskaja et al., 2017). e Western blot depicting Denr and actin expression in fetal HSPCs after electroporation with Cas9 LacZ guide DNA (control) and Cas9 guide designed to disrupt expression of Denr. f Median fluorescence intensity (MFI) of myeloid (CD11b) (left) and lymphoid (CD19) (right) markers in wild-type (WT) and Denr knock-out (KO) fetal HSPCs cells after 7 and 14 days of culture. The bar graph presents mean ± SD in two or three biological replicates. ****p < 0.0001 (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)