Figure 4.
Late loss of AnkG causes prolonged nodal destabilization compared with early loss of AnkG. A–I, SN fibers from 1, 3, and 10 mpi age-matched Ankfx/− and SLICK-H-CreER;Ankfx/− littermate mice were immunostained with antibodies against AnkG (blue) and Caspr (green) in combination with any of the following proteins: NF186 (A–C), βIV Spec (D–F), or pan-NaV (G–I; red). J–L, Graphs representing intensity quantification of NF186, βIV Spec, or pan-NaV, respectively, in the SN nodal area of SLICK-H-CreER;Ankfx/− (red bars) standardized to the same age control values from 1, 3, and 10 mpi Ankfx/− mice (black bars; n = 3 mice/genotype; 120–130 nodes/animal; unpaired, two-tailed Student's t test). M–U, Immunostaining of SC sections from 1, 3, and 10 mpi age-matched Ankfx/− and SLICK-H-CreER;Ankfx/− littermate mice with antibodies against AnkG (blue) and Caspr (green) in combination with any of the following proteins: NF186 (M–O), βIV Spec (P–R), or pan-NaV (S–U; red). V, W, X, Graphs representing intensity quantification of NF186, βIV Spec, or pan-NaV, respectively, in the SC nodal area of SLICK-H-CreER;Ankfx/− (red bars) standardized to the same age control values from Ankfx/− mice (black bars; n = 3 mice/genotype; 120–130 nodes/animal; unpaired, two-tailed Student's t test). Y, Z, Percentage of normal and patchy nodes in SNs and SCs, respectively, with the remaining NF186, pan-NaV, and βIV Spec at 3 and 10 mpi in SLICK-H-CreER;Ankfx/− mice compared with Ankfx/− controls (n = 3 mice/genotype; 120–130 nodes/animal; comparison of normal nodes performed by two-way ANOVA with Bonferroni's post hoc analysis). Z′, Immunoblot analysis of SC lysates from 3 mpi Ankfx/− and SLICK-H-CreER;Ankfx/− mice with antibodies against NF186, pan-NaV, βIV Spec, MBP, and tubulin (Tub; as a loading control). Arrowheads mark the nodes without AnkG in the SLICK-H-CreER;Ankfx/− mice. Scale bar, 2 μm. All data are represented as the mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.