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. 2016 Jan 18;7:10058. doi: 10.1038/ncomms10058

Figure 4. Rescue of axonal growth by kinesin mutants.

Figure 4

(a) In vitro expression of mutant TUBB3 and KIF5B/KIF21A in mouse cortex neuron was detected by immunofluorescence via TUBB3-fused V5 tag or kinesin-fused HA tag. Cells were also stained for axonal marker, tau-1. Scale bar, 5 μm. (b) Effects of R262H and R262A TUBB3, D279R KIF5B and D325R KIF21A overexpressions on axon length. Lengths of 110–200 axons were measured. Error bars indicate s.e.m. *, ** and NS indicate P<0.05, P<0.01 and P≥0.05, respectively (ANOVA followed by post hoc pairwise Wilcoxon–Mann–Whitney tests). (c) Commissural axons in P3 mouse brain, transfected with (from above to below) WT TUBB3, R262A TUBB3 alone and R262A TUBB3 plus D325R KIF21A. Commissural axons were stained by anti-GFP antibody reacting with EYFP reporter. (d) Commissural axons were stained by antibody labelling V5 tag of recombinant TUBB3. (c,d) The broken line and red arrowheads indicate the midline and the tip of the axon, respectively. Scale bar, 500 μm. (e) Horizontal distances between the midline and the tip of the axon bundle, averaged from four to nine mouse brains (mean±s.e.m.). *, *** and NS indicate P<0.05, P<0.001 and P≥0.05, respectively (ANOVA followed by post hoc Tukey–Kramer tests). (b,e) Single-molecule motility of the equivalent combination, data for which are shown in Fig. 3 and Supplementary Table 3, is indicated in blue (+/−) below the graph. NS, not significant.