Figure 6: Sema3A enhances PlexA4 transport in axons and in dendrites.

(a) Sema3A decreased the levels of PlexA4 immunostaining in distal axon and then increased the levels in dendrites. Magnified images of boxed areas in middle panels are shown in left panels. Relative mean immunostaining intensity in distal axon, proximal axon, soma and dendrites per neuron according to the counter staining of Tau1 (axon) and MAP2 (dendrites) is shown in b (n=60 neurons from three independent cultures). (c) Sema3A enhances fluorescence recovery of the ratio of PlexA4-EGFP to mcherry within the photobleached area of hippocampal axon, but not dendrite, from 1 to 5 min after Sema3A stimulation. Mean fluorescence recovery at 200–250 s after photobleaching is shown in d (n=9–14 neurons from more than three independent cultures). (e) Sema3A enhances fluorescence recovery of PlexA4-EGFP in the photobleached area of hippocampal dendrite, but not axon, from 13 to 17 min after Sema3A stimulation. Mean fluorescence recovery at 200–250 s after photobleaching is shown in f (n=12–16 neurons from more than three independent cultures). Scale bars in a, 5 μm (left panel); 20 μm (right panel); (c,e), 5 μm. ^†P<0.01 by ANOVA compared with 0 min. *P<0.05, **P<0.01 by t-test compared with vehicle control. Data are presented as mean±s.e.m.