Mitochondrial depolarization is a consequence of autophagosome formation in reticulocytes. Immunofluorescence of reticulocyte-enriched blood from phlebotomized Atg7^+/+ and Atg7^−/− transplant recipients, cultured for 3 days, and doubly stained with MTG and TMRM. The mean immunofluorescence intensity per cell of all cells with mitochondria in a representative field was quantified in the red and green channels, and the ratio of the signals was determined. The higher the ratio, the greater the average mitochondrial polarization per cell. The proportion of cells within each range is represented by a chart below each picture (legend at the bottom). Approximately 50 to 100 cells were quantified per sample (field). Note that mitochondria in the Atg7^−/− reticulocytes remain polarized, whereas mitochondria in Atg7^+/+ reticulocytes do not. No quantification is provided for Atg7^+/+ reticulocytes on day 3 due to an insufficient number of cells with mitochondria for analysis. The scale is indicated at the bottom.