Fig 9. Quantitative evaluation of membrane targeting of NaRs.

(A) Expression of eYFP-labeled I₁K₆NaR in the cultured cortical neuron. (i) Integrated confocal image of the neuron (Z-stack projection of 12 slices at every 0.46 μm in depth). (ii) eYFP image of a single slice shown in the dot-lined square in (i). (iii) Image of the fluorescent WGA of the same region. (iv) Merge of (ii) and (iii), indicating the expression of I₁K₆NaR in the neuronal membrane. (B) Profiling of the fluorescent intensity of an I₁K₆NaR-expressing neuron. The pixel values along the horizontal/vertical axes, which were delineated to pass the centroid of the cell (yellow), were plotted as functions of distance: eYFP fluorescence (green) and WGA fluorescence (magenta). The overlapping index was 0.68. (C) Profiling of the fluorescent intensity for a KR2-expressing neuron. The overlapping index was -0.21. (D) Comparison of the overlapping index between I₁K₆NaR (left) and KR2 (right). ***, P < 0.0005, Mann-Whitney U-test. Scales, 20 μm for (A) and 10 μm for (B) and (C).