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. 2012 Mar 19;196(6):727–742. doi: 10.1083/jcb.201107096

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© 2012 Ryan et al.

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Figure 9. — Dystonin-a2–MAP1B interaction maintains Ac–α-tubulin and promotes flux through the secretory pathway. (A) Western blot analysis of 293T cell lysate shows that MAP1B overexpression maintains Ac–α-tubulin after dystonin-a2 depletion relative to empty vector (EV) control. (B) MAP1B overexpression prevents Golgi fragmentation after dystonin-a2 depletion in 293Ts. Arrows point to intact Golgi in an siA2-treated cell that is positive for MAP1B-GFP expression. Arrowheads point to fragmented Golgi in a neighboring cell that was not transfected with the MAP1B-GFP construct. (C) MAP1B overexpression promotes VSVG trafficking to the Golgi and PM after dystonin-a2 depletion relative to empty vector control. Scramble siRNA-treated cells (top and A) are labeled C. The arrowhead identifies aberrantly localized VSVG molecules, whereas arrows identify normal VSVG localization. (D and E) Quantification of MAP1B impact on VSVG accumulation in the Golgi (D) and at the PM (E; ANOVA posthoc Tukey; n = 3). (F) MAP1B antigenic labeling of WT primary sensory neurons shows a perinuclear accumulation that is lost in dt^27J neurons (arrows). (G) MAP1B expression restores GLuc flux through the secretory pathway (time = 1 h) in P15 dt^27J sensory neurons (ANOVA posthoc Tukey; n = 6). Error bars show means ± SEM. Cont, control; Tub, tubulin. **, P < 0.01. Bars, 10 µm.