Fig. 4
Active immunization does not affect pTDP-43 levels and TDP-43 cytoplasmic-to-nuclear distribution in rNLS8 mice. Cerebral TDP-43 pathology was analyzed by ELISA, immunohistochemistry, and immunofluorescence stainings in vaccinated rNLS8 mice. A Phospho-TDP-43 (pS409/410) levels were measured in RIPA and urea fractions from neocortex. Only monogenic animals showed statistically significant differences in RIPA lysates (ctrl PBS: p = 7.63*10–5, ctrl VAX: p = 7.63*10–5) and urea lysates (ctrl PBS: p = 7.70*10–5, ctrl VAX: p = 7.70*10–5) when compared to TDP-PBS group by pairwise Wilcoxon test with Benjamini–Hochberg correction. Mean values are plotted as black dots. B Immunohistochemical staining in the hippocampal region using an antibody against pS409/410 TDP-43. Representative images for each group are shown. Magnified insets are displayed on the top right corner and their position in the overview is depicted by dashed rectangles. Arrows in the inset denote characteristic pTDP-43 inclusions in the CA3 region. Scale bar overview = 500 µm, scale bar inset = 50 µm. C: Immunofluorescence staining of total TDP-43 (human + mouse) in the frontal neocortex. Representative images for each group are depicted in the left panel (Scale bar = 20 µm). Note that control PBS images were taken at a higher gain to compensate for the lower levels of endogenous TDP-43. Right panel shows quantification of the ratios of the cytoplasmic-to-nuclear TDP-43 intensities (after normalization to the ctrl PBS group). Mean values are plotted as black dots. Pairwise t-test with Benjamini–Hochberg correction indicated a statistically significant difference between monogenic ctrl PBS and bigenic TDP-PBS animals (p = 3.32*10–5). Control (ctrl) PBS n = 6, TDP-PBS n = 6, TDP5 + 7 n = 5, TDP8 + 9 n = 6, TDP10 + 12 n = 5, and TDP15 n = 6