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. 2010 Oct 4;191(1):211–223. doi: 10.1083/jcb.201006039

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© 2010 Xiong et al.

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Figure 7. — Injury induces an increase in Wnd protein in axons concomitant with a decrease in Hiw. (A) Western blots from nerve cords with attached segmental nerves to detect endogenous Wnd protein levels (20 VNCs per lane) before and after injury. (B) Quantification of changes in Wnd protein level based on Wnd/tubulin ratio from five independent experiments. (C1 and C2) GFP-Wnd^KD particles in nerve cords and segmental nerves before and 4 h after injury. UAS–GFP-wnd^KD is expressed in motoneurons by OK6-Gal4. The cartoon shows the anatomy of nerve cord and segmental nerves, with the blue rectangles indicating the sites shown. (C1) Nerve cords of uninjured and injured animals. Injury induces dramatic increase of GFP-Wnd^KD intensity in the axons projecting from motoneuron cell bodies (yellow arrows). (C2) Segmental nerves proximal to injury site are shown in higher magnification costained with HRP to label the nerve membrane. (D and E) The injured segmental nerves contain a higher mean GFP intensity (D) and a higher density of GFP-Wnd^KD particles (E). Quantification is described in Materials and methods. (F1 and F2) GFP-Hiw (driven by OK6-Gal4) in nerve cords and segmental nerves in uninjured and injured (4 h) animals. (F1) Injury induces overall decrease of GFP-Hiw in indicated axons (yellow arrows). (F2) GFP-Hiw localizes to particles (white arrows) in uninjured axons, which are dramatically decreased in number within 4 h after injury. (G) Measurement of GFP-Hiw particle density in segmental nerves. P > 0.05 was not significant. *, P < 0.05. Error bars indicate mean ± SEM. Bars, 25 µm.