ChAc red cells show accumulation of multivesicular bodies and delayed mitochondrial clearance. (A) ChAc red cells of normal shape and spiculated acanthocytes (open arrowhead) are both found in peripheral blood of ChAc patients (upper left; bar, 2 μm). Ring-shaped membrane structures (solid arrowhead) are found in both acanthocytes (upper center and right panel; bar, 1 and 0.5 μm, respectively) and in ChAc red cells of normal shape (lower right panel; bar, 0.5 μm); remnants of individual and paired double membrane (lower left panel, higher original magnification in inset; bar, 0.5 and 0.2 μm, respectively) are also visible. In ChAc red cell, multivesicular bodies are found close to the cell membrane (lower center panel; higher original magnification in upper inset; bar, 0.5 and 0.2 μm, respectively) and may be in the process of being extruded from the cell (lower center panel, lower inset; bar, 0.5 μm). (B) Ulk1 and Atg7 were individually immunoprecipitated from red cell cytosol of healthy (Ctrl, control) and ChAc subjects and then subjected to immunoblot with anti-chorein antibody. The experimental results shown are representative of 6 similar experiments, each from an individual ChAc subject. IgG was used as loading control. Data are shown as means ± SD (n = 6; *P < .01 compared with ChAc subjects). (C) (Left) Flow cytometry of reticulocyte-enriched red cell fractions from healthy (Crtl, control) and ChAc subjects intravitally stained with MitoTrack. One representative of 3 independent experiments with similar results. (Right) Data are shown as means ± SD (n = 3; *P < .05 compared with healthy fractioned red cells).