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. 2004 Aug 16;166(4):571–578. doi: 10.1083/jcb.200312155

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Copyright © 2004, The Rockefeller University Press

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Figure 1. — A point mutation inhibits sodium channel clustering at the AIS. (A, left) Cell surface distribution of CD4-Na_v1.2 II–III in transfected hippocampal neurons. CD4-Na_v1.2 II–III was immunodetected with an anti-CD4 antibody (green) before permeabilization and the somatodendritic domain was subsequently identified by MAP2 staining (red). CD4-Na_v1.2 II–III was restricted at the AIS (arrow). (A, right) CD4-Na_v1.2 II–III was resistant to Triton X-100 extraction before cell fixation and colocalized with ankyrin G. (B) Cell surface distribution of the indicated CD4-Na_v1.2 II–III mutants. (C) Schematic representation of mutations within the AIS motif of Na_v1.2. (D) Histogram of the cell surface distribution of CD4-Na_v1.2 II–III mutants. For each mutant, the percentage of transfected CD4-positive hippocampal neurons were classified into three categories: CD4 staining concentrated at the AIS (hatched), localized on axons (gray shading), and uniformly distributed at the cell surface of somatodendritc and axonal domains (black) taking as 100% the total population of transfected neurons. Data are means ±SD from three to five different experiments. n denotes the total number of cells analyzed for each mutation. (E) In parallel, the resistance of mutated proteins to Triton X-100 extraction before cell fixation was determined. Bars, 20 μm.