Fig. 5: Detection of lysosome-mitochondria contact sites in vivo.
a Representative images (Z-projections from a complete Z-stack) of Mouse Primary Cortical Neurons (MPCNs) isolated from wt mice, stained with an anti β-TubIII (647 nm) and Hoechst (405 nm), with SPLICSS- P2ALY-MT probe (488 nm). Scale bar 25 μm. b Quantification of the Ly-Mt contacts was performed from the 3D rendering of a complete Z-stack. Data is shown as mean ± SEM dots/cell (left) or dots in axons/total dots (SPLICSS-P2ALY-MT 20.19 ± 2.17 n = 34, soma; 0.23 ± 0.035 n = 34, axon, n = cells examined over 4 independent experiments). c Live imaging of Ly-mt contact sites in zebrafish RB neurons, representative confocal images of SPLICSS-P2ALY-MT in RB neurons of 24 hpf s1102t:GAL4 living embryos injected with the pT2-DsRed-UAS-SPLICSS-P2ALY-MT construct. Rostral is on the right, dorsal on the top. d Quantification of the Ly-Mt contacts was performed from the 3D rendering of a complete Z-stack, mean ± SEM: SPLICSS-P2ALY-MT 24.53 ± 2.607 n = 15 cells over at least 3 independent injections. e Live imaging and (f) quantification of Ly-mt contact sites in D. melanogaster wing sensory neurons (n = 36 wings) from 2-day old flies (Mean ± SEM: SPLICSS- P2ALY-MT 18.81 ± 0.6471 n = 36). Dashed line: boundaries of the neuronal tract. The soma and the axon are indicated with red arrowheads and white arrows, respectively. Data points represent individual SPLICS puncta from 50 μm of axons. “Parts of the figure were drawn by using pictures from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/).” Source data are provided as a Source Data file.
