Video 5

Movie S5. Three-dimensional reconstruction of confocal stacks of the optic nerve head from a mouse that received an adenoassociated virus 2 (AAV2)::mtDNA (Mito) EGFPmCherry intravitreal injection shows that mCherry-positive/EGFP-negative (mCherry-ONLY) signal colocalizes with lysosomal-associated membrane protein 1 (Lamp1). These are the same data presented in Fig. 2C. Stacks of confocal images (Z = 0.4 μm) were rendered as 3D reconstructions using Imaris software (Bitplane, Inc.). DAPI is shown in blue, EGFP is shown in green, mCherry is shown in red, and Lamp1 is shown in white. EGFP and mCherry are initially shown separately and then together to emphasize the presence of acidified mitochondria. mCherry-ONLY signal (magenta) is overlaid on the EGFP-mCherry signal in flashes, and then shown by itself. White channel (Lamp1) signal is flashed over the mCherry-ONLY signal. Then, the mCherry-ONLY signal is phased out. EGFP and mCherry signals are phased back in with the white Lamp1 signal.