In late-phase erythropoiesis, ChAc erythroblasts exhibit impaired autophagy with delayed clearance of lysosomes. (A) Western blot analysis of p62 and LAMP1 in day 14 erythroblasts cultured from healthy (Crtl) and ChAc subjects. Catalase was used as protein loading control. Densitometric analysis of the immunoblot bands are presented at right with data as means ± SD (n = 6; *P < .01 compared with controls). (B) (Upper) LAMP1 immunostained cytospin preparations of day 14 erythroblasts from healthy (Crtl) and ChAc subjects counterstained with DAPI. (Lower) The puncta mean fluorescence was measured using Image J software. Data are presented as means ± SD (n = 4); *P < .05 compared with healthy cells. (C) (Upper) Flow cytometry of LysoTracker-stained day 14 erythroblasts from healthy (Crtl) and ChAc subjects (late-E, late-erythroblasts; Int-E, intermediate erythroblasts). One representative experiment of 4 separate experiments with similar results. (Lower) Data are shown as means ± SD (n = 4; *P < .05 compared with healthy cells).