In vivo induction of mitophagy through an optogenetic bimodular system. a Zebrafish embryos were microinjected with a Venus-iLID-ActA overexpressing plasmid, then fixed 48-hpf and whole mount immuno-stained for Tom20 (red). Nuclei were counterstained with DAPI (blue). Inset: ×4 magnification. Scale bar: 10 μm. b Venus-iLID-ActA/AMBRA1-RFP-sspB microinjected zebrafish embryos were illuminated with blue light and then kept in the dark for 3 min, mimicking the experiment shown in Fig. 3a. Double-positive muscle fibers were photographed in order to analyze Venus-iLID-ActA/AMBRA1-RFP-sspB dynamic interactions. PCC and MOC of the red over the green signal were quantified in three independent experiments. Inset: ×6 magnification. Scale bar: 10 μm. Data shown: mean ± S.E.M. Hypothesis test: ANOVA test. *p < 5 × 10−2. **p < 10−2. c Venus-iLID-ActA/AMBRA1-RFP-sspB or Venus-iLID-ActA/RFP-sspB (negative control) double positive zebrafish embryos were illuminated for 8 h with a pulsed (2 s light/2 min dark) blue light; muscle fibers were analyzed. Upon light stimulation, round-shaped mitochondria (arrowheads) were clearly visible in AMBRA1-RFP-sspB but not RFP-sspB positive fibers. Images are the sum of three frames Z-stack. Stars indicate nuclei. Graphs show the overall reduction per single fiber of the Venus-iLID-ActA signal intensity corrected for the background in three independent experiments. Inset: ×8 magnification. Scale bar: 10 μm. Data shown: mean ± S.E.M. Hypothesis test: ANOVA test. *p < 5 × 10−2. Source data are provided as a Source Data file
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