EHBP1L1 cooperates with Rab10, Bin1, and dynamin to promote erythroblast enucleation. (A-C,F) Lin⁻Ter119⁻CD117⁺ erythroid progenitors from ICR embryos (A-C) and embryos with the indicated genotypes (F) were differentiated as described in the “Materials and methods” section. The differentiated erythroblasts were subjected to immunofluorescence staining for the indicated proteins. The white arrows indicate representative colocalization events. Nuclei (blue) were stained with 4′,6-diamidino-2-phenylindole (DAPI). Bars: 2 μm. (D-E,G-J) MEDEP-BRC5 cells with shRNA-mediated knockdown (KD) of Ehbp1l1, Rab10, or Bin1 (D-E,G-H) or with dynamin inhibitor treatment (I-J) were differentiated and analyzed by flow cytometry. The cells were treated with the dynamin inhibitors (30 μM) for the last 24 hours of differentiation (I-J). Representative flow cytometry plots of PI⁻Ter119^high cells are shown. The percentage of Hoechst 33342⁻ cells among PI⁻Ter119^high cells is shown in a bar-dot plot (triplicates; E,H,J). The data are representative of more than 3 independent experiments. The data are presented as means ± standard deviation (SDs). ∗∗∗∗P < .0001 (1-way ANOVA in panel E,J; unpaired t test with Welch’s correction in panel H).