Skip to main content
. 2019 May 29;39(22):4238–4251. doi: 10.1523/JNEUROSCI.2771-18.2019

Figure 5.

Figure 5.

Nav channel expression is unaltered by WT or MAPM Nav1.6 transfection. A, Representative TIRF images of a DIV 7 rHN expressing Nav1.6-LoopBAD-mCherry WT in the absence (top) or presence (bottom) of Nav1.6-GFP MAPM. mCherry fluorescence is shown in the first column (yellow), surface labeling with SA-CF640 in the second column (magenta), and GFP fluorescence in the third column (cyan). Scale bars, 10 μm. B, Average values for SA-CF640 fluorescence of WT Nav1.6 in the AIS (top) or somatic (bottom) compartments. Error bars indicate mean ± SEM. The difference in AIS labeling was not significant. For the mean SA-CF640 AIS signal: 519.8 ± 106.9 AU for and 364.2 ± 45.8 AU for WT-LoopBad-mCherry alone and WT-LoopBad-mCherry + MAPM-GFP, respectively. The decreased surface expression of the WT on the soma in the presence of MAPM was significant. For SA-CF640 soma signal: 121.3 ± 23.5 AU and 68.1 ± 10.0 AU for the WT-LoopBad-mCherry alone and WT-LoopBad-mCherry + MAPM-GFP, respectively. C, Representative TIRF images of DIV 7 rHN expressing Nav1.6-LoopBAD-GFP MAPM in absence (top) or presence (bottom) of Nav1.6-mCherry WT. The GFP fluorescence is shown in the first column (cyan), surface labeling with SA-CF640 in the second (magenta), and the mCherry fluorescence in the third column (yellow). Scale bars, 10 μm. D, Average values of SA-CF640 fluorescence in the AIS (top) or somatic (bottom) compartments. Error bars indicate mean ± SEM. The differences in surface labeling in both the AIS and soma were significant. For the SA-CF640 AIS signal: 58.2 ± 8.8 AU and 31.5 ± 4.0 AU for MAPM Nav1.6-LoopBad-GFP alone and MAPM Nav1.6-LoopBad-GFP + WT Nav1.6-mCherry, respectively. For the SA-CF640 soma signal: 36.4 ± 4.9 AU and 24.2 ± 3.3 AU for MAPM Nav1.6-LoopBad-GFP alone and MAPM Nav1.6-LoopBad-GFP + WT Nav1.6-mCherry, respectively. E, Representative AIS maximum intensity Z projections from DIV 7 rHN cultures either untransfected, that is, endogenous (left), transfected with WT Nav1.6-GFP (middle) or MAPM Nav1.6-GFP (right). GFP (cyan) and PanNav immunolabeling staining (magenta) are shown. Scale bars, 5 μm. F, Average values of AIS PanNav staining of untransfected (endogenous), WT, or MAPM transfected cells. Error bars indicate mean ± SEM. *p ≤ 0.05, **p ≤ 0.01.