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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: RNA binding proteins co-localize with small tau inclusions in tauopathy

Fig. 4

RNA binding proteins show significant colocalization with diffuse phospho-tau but not NFTs in the rTg4510 cortex. (a) Immunohistochemical analysis of rTg4510 tissue (n = 3) has also revealed a significant colocalization in the cortex between the RBPs DDX6, PABP, HNRNPA0, and eIF2a (red) with pathological phospho-tau stained using the CP13 antibody (green). However, the RBP and splicing factor U2AF2 does not show significant correlation. To the right of each merged image is a scatterplot of the pixel intensities for each pixel of the image in the red channel vs. the green (Pearson correlation coefficients r = 0.773 for DDX6, 0.791 for eIF2α, 0.325 for HNRNPA0, 0.798 for PABP, and − 0.14 for U2AF2). This colocalization is greatly reduced and/or completely lost as tau aggregates into large NFTs which are brightly fluorescent and fill the cell bodies of neurons (b) (r = 0.069 for DDX6, 0.372 for eIF2α, 0.481 for PABP, − 0.03 for HNRNPA0, and − 0.009 for U2AF2). c Staining of wild-type C57Bl/6 mice also indicates that HNRNPA0 is predominantly nuclear in healthy animals, while the rTg4510 staining shows significant cytoplasmic localization of HNRNPA0 (a, b). (d) Negative controls IHC using rabbit and mouse normal IgG indicates that there is no off target staining or fluorescence in our tissues. e Pearson coefficients of correlation between CP13 positive tau with RBPs DDX6, eIF2α, HNRNPA0, PABP, and U2AF2 are graphed for individual neurons using ImageJ. For all cases except U2AF2, neurons show heterogeneity in colocalization between phospho-tau and the RBPs stained, from no colocalization to fully overlapping reactivity patterns in individual neurons. The percent of neurons with r > 0.3 is graphed in (f) as the percentage of neurons showing moderate to strong correlations between green:red intensity (DDX6 = 36% of neurons; eIF2α = 54% of neurons; HNRNPA0 = 35% of neurons; PABP = 33% of neurons; U2AF2 = 0% of neurons)

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