Somatic mitophagy preferentially removes axon-derived mitochondria after reperfusion. (A and B) Primary cultured cortical neurons were previously transfected with MitoDendra2 and tagBFP-LC3B by electroporation. After photo-conversion of MitoDendra2 in axons, the cultured neurons were subjected to 20 min of OGD and 20 min of reperfusion (O-R) as shown in A in DIV8. (B) Images show representative examples from three independent experiments. Orthogonal view and line-scan analysis of indicated regions are presented in the right panel. (C–E) Primary cultured cortical neurons expressing mCherry-LC3B were stained with MTG (green) in two different patterns as shown in C in DIV8. After 20 min of OGD followed by 20 min of reperfusion, fluorescent images were captured by confocal microscopy. (D) Images show representative examples from three independent experiments. Orthogonal view and line-scan analysis of indicated regions are presented in the right panel. (E) Columns represent the number of mitophagosomes (upper panel) and autophagosomes (lower panel) in neuronal soma. n = 17–22 cells from three independent experiments for each group. The data are expressed as means ± SEM. Statistical comparisons were performed with one-way ANOVA with Sidak’s multiple-comparisons test. **P < 0.01, ***P < 0.001 versus the indicated group. (F–G) Neurons expressing MitoQC were stained with MTDR (cyan) and then subjected to 20 min of OGD and 20 min of reperfusion (O-R) as shown in F in DIV8. (G) Images show representative examples from three independent experiments. Orthogonal view and line-scan analysis of indicated regions are presented in the right panel. Scale bar, 1 µm. A.U., arbitrary units; Ctrl, control; Mito, mitochondria.