Fig. 3

Microtubules in nudc axon terminals show signs of enhanced stabilization (A and B) Staining of Tubb3 in wild type and nudc terminals. (C) Quantification of Tubb3 fluorescence intensity shows no difference in nudc terminals (ANOVA). (D and E) Labeling of acetylated tubulin in wild type and nudc terminals. White arrow in (E’) highlights the net-like organization of acetylated tubulin staining in mutants (inset). (F) Quantification of acetylated tubulin fluorescence intensity shows no difference in nudc terminals though the organization is altered (ANOVA). (G and H) Acetylated tubulin labeling in axon terminals of two unrelated mutants, p150 and actr10, show no change in microtubule structure despite changes in the axon terminal structure. (I and J) Wild type (I) and nudc mutant (J) axon terminal showing EB3-GFP comets. (K) Axon terminal comet number is reduced in nudc mutants (ANOVA). (L–N) Dendra2-labeled α-tubulin in an axon terminal of the pLL. Prior to conversion, (L) no red fluorescence is detected (shown in magenta). Post-conversion, strong labeling of converted Dendra2-α-tubulin is observed (M). Converted Dendra2-α-tubulin decay over 30 min post-conversion shown in N. (O) Decay curve of converted Dendra2-α-tubulin from time-lapse imaging post-conversion demonstrates a shift in the half-life in nudc mutants (mixed effects model; p = 0.0033). Data are expressed as mean ± S.E.M. Scale bar = 10 μm. Sample sizes indicated on graphs.