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Fig. 3 | Molecular Brain

Fig. 3

From: Schizophrenia-associated dysbindin modulates axonal mitochondrial movement in cooperation with p150glued

Fig. 3

Dysbindin regulates axonal mitochondrial motility in cooperation with p150glued. a Representative kymographs of mitochondrial movement in the proximal axons of DIV 10–12 primary cortical neurons transfected as indicated. b Quantitative analysis of the motile fraction of mitochondria in the proximal axons (n = 32 axons for control (CTL) shRNA, n = 33 axons for p150glued shRNA, n = 14 axons for Dysbindin shRNA, and n = 24 axons for p150glued shRNA + Dysbindin shRNA). c Quantification of mitochondrial density (the number of mitochondria per 100 μm) in the distal axons of DIV 10–12 primary cortical neurons (n = 26 axons for CTL shRNA, n = 12 axons for p150glued shRNA, n = 9 axons for Dysbindin shRNA, and n = 17 axons for p150glued shRNA + Dysbindin shRNA). d Representative kymographs of mitochondrial movement in the proximal axons of DIV 10–12 primary cortical neurons transfected as indicated. e Quantitative analysis of the motile fraction of mitochondria in the proximal axons (n = 34 axons for CTL shRNA, n = 30 axons for p150glued shRNA, n = 32 axons for mDysbindin, and n = 33 axons for p150glued shRNA + mDysbindin). f Quantification of mitochondrial density (the number of mitochondria per 100 μm) in the distal axons of DIV 10–12 primary cortical neurons (n = 49 axons for CTL shRNA, n = 27 axons for p150glued shRNA, n = 34 axons for mDysbindin, and n = 30 axons for p150glued shRNA + mDysbindin). g Schematic diagrams of deletion mutants of dysbindin protein used for the co-immunoprecipitation analysis. aa, amino acids. h Co-immunoprecipitation of p150glued with dysbindin fragments. Lysates from HEK293 cells transfected with FLAG-p150glued and indicated GFP-hDysbindin fragments were immunoprecipitated with anti-FLAG antibody. Immunoprecipitates were analyzed by western blotting with anti-GFP and anti-FLAG antibodies. i Representative kymographs of mitochondrial movement in the proximal axons of DIV 10–12 primary cortical neurons transfected as indicated. j Quantitative analysis of the motile fraction of mitochondria in the proximal axons (n = 15 axons for CTL shRNA, n = 18 axons for Dysbindin shRNA + Control vector (pFLAG-CMV2), n = 20 axons for Dysbindin shRNA + mDysbindin WT, n = 24 axons for Dysbindin shRNA + mDysbindin 1–87 aa, and n = 9 axons for Dysbindin shRNA + mDysbindin 88–352 aa). Scale bars represent 20 μm. All results are presented as the mean \(\pm\) SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 from one-way ANOVA with Bonferroni’s multiple comparison test

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