FIGURE 3.

Analysis of ARD-dependent localization of Ank2-L and Ank2-XL in second instar ank2^ΔL mutants. (A–G) Analysis of Ank2-L (green) and Ank2-XL (magenta) localization at muscle 4 NMJs (HRP, white). (A) In control animals (P[ank2_wt]/+; ank2^ΔL/ank2^null) Ank2-L and Ank2-XL level are restored compared to ank2^ΔL/ank2^null mutants. (B) In ank2^ΔL/ank2^null mutant animals Ank2-L is absent and levels of Ank2-XL are severely reduced at the NMJ and also in the axon. (C–G) Analysis of the AnkD manipulations (P[ank2_ΔAnkDx]/+; ank2^ΔL/ank2^null) reveals alterations of Ank2-L and Ank2-XL similar to those in third instar mutant larvae. The ΔAnkD1 rescue construct almost completely restores Ank2-L level. All deletion mutations significantly reduce Ank2-XL distribution at the NMJ and in axons. (H) Quantification of Ank2-L level at the NMJ, within distal boutons and in the axon (n = 11–12 muscle 4 NMJs, three animals/genotype). (I) Quantification of Ank2-XL level at the NMJ, in distal boutons and in the axon (n = 11–12 muscle 4 NMJs, three animals/genotype). Scale bars in (A) apply to (A–G) and represent 10 μm. Error bars indicate SEM; ^∗∗p < 0.01, ^∗∗∗p < 0.001 (ANOVA); black asterisks represent comparison to controls; red asterisks represent comparisons to ank2^ΔL mutants.