Effect of dominant-negative mutants of μ subunits on somatodendritic sorting of NiV-F.

<p>(A–D) Rat hippocampal neurons were co-transfected with plasmids encoding NiV-F-GFP and wild-type (WT) or dominant-negative (DN) mutants of HA-tagged μ subunits (A-site mutants of μ1A, μ2, μ3A and μ4, and B-site mutant of μ4, as labeled on top of images). The effects of μ1A, μ2, μ3A and μ4 constructs are shown in panels A, B, C and D, respectively. Cells were immunostained with mouse anti-HA, rabbit anti-MAP2 and goat anti-ANK-G, and imaged as indicated in the legend to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004107#ppat-1004107-g001" target="_blank">Figure 1</a>. Grayscale images shown in all panels correspond to NiV-GFP fluorescence. The anti-HA, anti-MAP2 and anti-ANK-G staining is shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004107#ppat.1004107.s002" target="_blank">Figure S2</a>. Two images corresponding to different effects on NiV-F-GFP sorting (representing ∼75 and 25% of the neuronal population) observed following expression of μ2 D174A/W419S are shown in (B). The AIS and axons are marked by cyan and red arrowheads, respectively. Scale bar: 20 µm. See <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004107#ppat-1004107-t001" target="_blank">Table 1</a> for polarity indexes.</p