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. 2006 Apr 24;173(2):265–277. doi: 10.1083/jcb.200511055

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Copyright © 2006, The Rockefeller University Press

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Figure 7. — Active JNK and S73-phosphorylated SCG10 colocalize in vesicular fractions. (A) Confocal sections of cortical neurons at 1 d in vitro immunostained for SCG10, PSCG10, and PJNK (green) and for β-tubulin (red) revealed conspicuously punctate immunoreactivity in the neurites. Corresponding staining in the absence of 1° antibody (−1°αr) is shown. Golgi (arrow) and growth cone–localized SCG10 (arrowheads) are indicated. The specificity of α-PSCG10 for immunostaining is shown in Fig. S1 (available at http://www.jcb.org/cgi/content/full/jcb.200511055/DC1). Bar, 10 μm. (B) To examine whether SCG10 and active JNK colocalized in vesicles, rat forebrain was fractionated. Homogenate (H) was centrifuged at 800 g to isolate intact cells and nuclei (P1). The remaining supernatant (S1) was centrifuged at 16,000 g to yield a pellet containing endosomal and Golgi fractions (P2) and supernatant (S3). The final pellet (P3) contains plasma membranes.