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Figure 5


Fig. 5. NCAM is partially recycled to the plasma membrane by Rab11-positive endosomes. (A) Cortical neurons isolated from E15.5 mice (NCAM+/–) were transiently transfected with NCAM140. B35 cells were transiently co-transfected with either NCAM140 or NCAM180 and GFP-Rab11. Endocytosis was induced for 1 hour by application of an NCAM-specific antibody. Cells were then fixed and processed for immunofluorescence analysis by visualization of NCAM with secondary Cy3-conjugated antibodies and in the case of cortical neurons with anti-Rab11-antibodies and secondary Alexa-Fluor-488-conjugated antibodies. (B) Quantification of colocalization of NCAM-positive vesicles with Rab5, Rab4, Rab11 or LysoTracker. Data are the mean ±s.e.m. of three independent experiments with at least 15 cells analysed in each experiment.





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