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. 2013 Aug 5;202(3):495–508. doi: 10.1083/jcb.201302078

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© 2013 Fu and Holzbaur

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 1. — JIP1 knockdown disrupts both anterograde and retrograde transport of APP-positive vesicles. (A) Representative images and line scans of APP-YFP intensity show that JIP1-knockdown DRGs contain fewer APP-positive vesicles in the axon than control DRGs. (B) JIP1 knockdown in DRGs significantly decreased the number of APP-positive vesicles in the axon. Control, 0.85 ± 0.08/µm; JIP1 siRNA, 0.36 ± 0.04/µm. (B–F) Data represent three independent experiments (n = 15–23 neurons). (C) Kymographs of APP-YFP motility in DRG transfected with siRNA against JIP1. Kymographs represent cumulative organelle movement (displacement on the x-axis) over time (y-axis). Arrested vesicles appear as vertical lines, whereas motile vesicles appear as diagonal lines toward either the right (anterograde) or left (retrograde). Bars: (horizontal) 5 µm; (vertical) 15 s. (D) JIP1 depletion significantly alters the directional distribution of APP-positive vesicles, causing decreases in the percentages of anterograde and retrograde vesicles and an increase in the percentage of arrested vesicles. Transport changes induced by JIP1 depletion are fully rescued by expression of a human JIP1 cDNA resistant to the siRNA. (E and F) JIP1 depletion significantly decreases mean run lengths and speeds of APP-positive vesicles in both anterograde and retrograde directions. Means represent only vesicles categorized as motile (i.e., anterograde or retrograde in D). Error bars show the mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001.