Figure 4.

Treatment with triciribine alters length of the Na_v1.6 immunofluorescence at the AIS but not fluorescent intensity of Na_v1.6. Representative images of the CA1 region of (A–C) DMSO and (D–F) triciribine treated slices, with (A,D) DAPI (gray), (B,E) NeuN (blue), and (C,F) AnkyrinG (red) and Na_v1.6 (green) merging. Scale bar in (F) is 10 µm. Zoom to AIS in (G–I) DMSO and (J–L) triciribine-treated slices with (G,J) Na_v1.6 (green), (H,K) AnkyrinG (red), and (I,L) merging in both channels, and the white bar indicates an analyzed 5 µm cross-section. Scale bar in (L) indicates 5 µm. Stratum oriens (so), stratum pyramidale (sp), and stratum radiatum (sr) are indicated. (M) Treatment with 20 µM triciribine for 2 h significantly increased the length of the Na_v1.6+ fluorescent intensity at the AIS, but (N) does not alter the fluorescent intensity of Na_v1.6 as measured by the cross-section of the AIS. Data shown are mean ± SEM and separated scatter graph with bars to illustrate biological replicates (n = 4 brain slices per condition) in panel (M). * p = 0.0104 as analyzed by a two-tailed nested t-test. Data were log normalized before analysis. Zeiss 63× oil immersion objective (1.4 NA) was used in all cases.