Defect in endolysosomal and autolysosomal formation and proteome remodeling in mDia2-deficient erythroid cells. (A) TEM studies of reticulocytes from mDia2^fl/flVav-Cre mice (bottom) and mDia2^fl/fl mice (top). An adjacent orthochromatic erythroblast (bottom right) with numerous organelles and vesicles is also seen. Bars represent 1 μm. Arrows in inset III indicate vesicles. Insets II and III show increased organelles compared to inset I. (B) Lineage-negative cells from the bone marrow of the indicated mice were cultured in erythropoietin-containing medium for 2 days. Quantitative analyses were performed of extracellular vesicles from the culture medium. *P < .05. (C) TEM analysis of a representative orthochromatic stage erythroblast from mDia2^fl/flVav-Cre mice. Images are representative of 10 randomly selected fields. (D) Volcano plot of a quantitative proteomic study. Proteins significantly upregulated (red) or downregulated (blue) in mDia2-deficient reticulocytes are shown. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of proteins upregulated or downregulated by >25% are shown. The number of proteins per group is indicated on the right. The experiments were repeated 3 times, and the data were obtained with combined individual analyses.