Defects of autophagosome-lysosome fusion in mDia2-deficient erythroid cells. (A) Lineage-negative cells from the bone marrow of the indicated mice were cultured in erythropoietin-containing medium for 2 days. Representative images of LC3B-stained orthochromatic erythroblasts are shown. Bars represent 2 μm. Images are representative of 10 randomly selected fields. The bottom panels illustrate a binucleated orthochromatic erythroblast. (B) Western blot analysis of the cells in panel A. (C) Lineage-negative cells from the bone marrow of the indicated mice were cultured in erythropoietin-containing medium for 2 days. Flow cytometric analysis was performed to determine GFP intensity on the Ter119⁺ DNA⁺ erythroblasts. (D) Western blot analysis of the cells in panel C for the detection of GFP-LC3 and LC3. (E) Lineage-negative cells from the indicated mice were transduced with Lenti-mCherry-EGFP-LC3. The cells were cultured in erythropoietin-containing medium for 2 days. Representative images of erythroblasts at different developmental stages based on the nuclear size were presented. Bars represent 10 μm. Images are representative of 10 randomly selected fields.