Posttranslational changes in TAR6/6 mice.

(A-D) Localization of tTDP-43, CTF-35 and hTDP-43 after cellular fractionation of 3 months old TAR6/6 mice. (A) Cell fractions of midbrain and cortex samples of 3 months old ntg, TAR6 and TAR6/6 mice were analyzed by Western blotting and probed with the indicated antibodies (GAPDH as cytoplasmic and HDAC3 as nuclear marker; long exp. = long time ECL exposure for 30 min). One representative example of 4 is shown (C: cytoplasm, N: nucleus). (B- D) Densitometric analysis of tTDP-43, CTF-35 and hTDP-43 levels in cell fractions (cyt: cytoplasm, nuc: nucleus). Nuclear tTDP-43 and CTF-35 levels of ntg and hTDP-43 of TAR6/6 were set as 100%. (E-H) Soluble and insoluble TDP-43 protein levels in brains of 3 months old TAR6/6 mice. Brain samples were separated into RIPA soluble (R) and RIPA insoluble / UREA fractions (U). Three representative examples of 5 are shown. Fractions were analyzed by Western blotting and probed with the indicated antibodies (long exp. = long time ECL exposure for 30 min). Coomassie Blue staining was performed as loading control for UREA fraction. (F-H) Densitometric analysis of tTDP-43, CTFs and hTDP-43 levels in RIPA and UREA fractions. For tTDP-43 and CTF evaluation, RIPA fraction of ntg was set as 100%, whereas for hTDP-43 analysis, RIPA sample of TAR6/6 was set as 100%. (B-D; F-H) Two-way ANOVA followed by Bonferroni‘s post-hoc test. Mean+SEM. *significances between genotypes, ^#significances between fractions. *p<0.05, **p<0.01, ***p<0.001.