thousand mitochondrial structures were analyzed per condition (= neurons at day 40 or day 100). For quantification, all samples within one experiment were stained simultaneously and imaged with identical settings. For analysis of mitochondrial trafficking studies, kymographs were created using the ‘Multiple Kymograph’ plugin in ImageJ software from time-lapse images. From these kymographs the movement of mitochondria was classified as retrograde (moved ≥10μm in the retrograde direction), anterograde (moved ≥10μm in the anterograde direction), or stationary (moved < 10μm during the duration of the 3 minutes video). The percent motility of mitochondria along the neural process (retrograde, anterograde, stationary) was calculated as a percentage of the total number of mitochondria imaged per neuronal process.
