Fig. 2

DISC1-SNPH complex regulates the mitochondrial movement in axons. a Reversal of DISC1 knockdown effect by SNPH knockdown. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 10–14. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 32 axons, 367 mitochondria for control shRNA, n = 30 axons, 385 mitochondria for mDISC1 shRNA, n = 32 axons, 387 mitochondria for mSNPH shRNA, and n = 34 axons, 397 mitochondria for mDISC1 shRNA and mSNPH shRNA). b Effect on dendritic mitochondrial movement. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 10–14. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 33 dendrites, 337 mitochondria for control shRNA, n = 31 dendrites, 392 mitochondria for mDISC1 shRNA, and n = 32 dendrites, 338 mitochondria for mSNPH shRNA). c Decreased axonal mitochondrial movement by SNPH. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 10–14. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 36 axons, 446 mitochondria for control, n = 34 axons, 451 mitochondria for FLAG-mDISC1, n = 33 axons, 428 mitochondria for mSNPH-Myc, and n = 37 axons, 464 mitochondria for FLAG-mDISC1 and mSNPH-Myc). d Effect of SNPH knockdown and DISC1 overexpression. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 11. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 34 axons, 415 mitochondria for mSNPH shRNA, and n = 33 axons, 432 mitochondria for FLAG-mDISC1 and mSNPH shRNA). e Enhanced mitochondrial anchoring by SNPH^∆481–495. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 10. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 31 axons, 398 mitochondria for control, n = 31 axons, 402 mitochondria for mSNPH-Myc, and n = 30 axons, 392 mitochondria for mSNPH^∆481–495-Myc). f Enhanced axonal mitochondrial anchoring by SNPH overexpression upon DISC1 knockdown. Primary cortical neurons were transfected as indicated followed by time-lapse imaging at DIV 10. i) Representative kymographs and ii) quantitative analyses of mitochondrial movement (n = 30 axons, 351 mitochondria for mSNPH-Myc, and n = 32 axons, 368 mitochondria for mDISC1 shRNA and mSNPH-Myc). Scale bars; 10 μm. Error bars are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, NS, not significant, by student’s t-test (Fig. 2d and f) or one-way ANOVA with Bonferroni’s multiple comparison test (Fig. 2a, b, c, and e)