Fig. 9: ER and DRP1 mark the site of pre-peroxisome formation. | Nature Communications

Fig. 9: ER and DRP1 mark the site of pre-peroxisome formation.

From: Quantitative optical nanoscopy of mitochondrial-derived vesicles in neurons classifies pre-peroxisomal and clearing organelles

Fig. 9

a Two-color STED image of ER (SNAP-ER, cyan) and mitochondria (Halo-OMM, magenta), representative of more than three independent experiments. ROI I shows detachment of a lateral protrusion at an ER contact-site (see Supplementary Movie 5). The schematic illustrates ER-mitochondria interactions during mitochondrial-vesicle release. b Two-color live STED image of ER (SNAP-ER, cyan) and mitochondria (Halo-OMM, magenta), representative of more than three independent experiments. ROI II shows detachment of a lateral protrusion at an ER contact-site (see Supplementary Movie 6). The schematic illustrates ER-mitochondria interactions during mitochondrial-vesicle release. c Three-color STED image of mitochondria, PEX16 and ER, representative of (e), showing MDS and mitochondrial protrusion with PEX16 enrichment, with ER tubules marking the pre-peroxisome formation site, confirmed by the line profile at position #1. d Three-color STED image of mitochondria, PEX14 and ER, representative of (f), showing a mitochondrial protrusion with PEX14 enrichment, with ER surrounding the protrusion, confirmed by the line profile at position #2. e Box plot of normalized mitochondrial protrusion frequency relative to proximity to ER and/or PEX16. Each data point represents the average number of protrusions per neuron; Nneurons = 9, Nprotrusions = 102. KS tests; *P < 0.05 (P = 0.03), ***P < 0.001 (P = 7.4 × 10−4; P = 7.4 × 10−4). f Box plot of normalized mitochondrial protrusion frequency relative to proximity to ER and/or PEX14. Each data point represents the average number of protrusions per neuron; Nneruons = 4, Nprotrusions = 69. KS tests; *P < 0.05. g Representative three-color STED image, relative to (h) of mitochondria (Halo-OMM, magenta), PEX14 (immunostaining, yellow), and DRP1 (immunostaining, cyan) showing DRP1 at PEX14+ mitochondrial-protrusion sites. h Box plots of normalized mitochondrial protrusion frequency relative to proximity to DRP1 and/or PEX14 (Nneurons = 10, Nprotrusions = 112) and DRP1 and/or ER (Nneurons = 5, Nprotrusions = 37). Each data point represents the average number of protrusions per neuron. KS tests; **P < 0.01 (P = 0.002; P = 0.008). i Sketch of mitochondria, ER, and DRP1 at pre-peroxisome formation site. For the box plots in (e, f, h), the center line represents the median, the box spans the interquartile range (IQR; 25th to 75th percentiles), and whiskers extend to 1.5× IQR.

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