(A) Hippocampal neurons were transfected with DsRed-mito, and time-lapse imaged to monitor mitochondrial movement within axons.
(B) The kymograph for the time-lapse images of (A).
(C, D) The image taken in the same region of the axon as (A) after fixation and staining with an anti-SNPH antibody immediately following time-lapse imaging. Scale bar: 10 μm. Arrows point to the mitochondria tagged with SNPH (yellow) and arrowheads indicate the SNPH-negative mitochondria (red). Note the correlation between the endogenous SNPH-tagged mitochondria in (C) and stationary mitochondria (vertical line) in (B).
(E) A complementary relationship between the mean percentages of the SNPH-tagged mitochondria (black, 1126/1727, 54 images) and the motile mitochondria (red, 120/305, 12 time-lapse images).
(F, G) Scatter plots with 90% confidence intervals show the percentages of the SNPH-tagged mitochondria (62±14%, 54 images)(F) and the stationary mitochondria within axons (62±15%, 12 time-lapse images)(G).