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Fig. 8 | Acta Neuropathologica Communications

Fig. 8

From: RBM45 associates with nuclear stress bodies and forms nuclear inclusions during chronic cellular stress and in neurodegenerative diseases

Fig. 8

RBM45 subcellular distribution and inclusion pathology in lumbar spinal cord. Immunohistochemistry for RBM45 with either SAFB or TDP-43 was performed in lumbar spinal cord sections from ALS and non-neurologic disease control subjects. a RBM45, SAFB, and TDP-43 immunoreactivity were measured in motor neurons and glial cells of non-neurologic disease control subjects. RBM45 exhibited strong nucleolar immunoreactivity in motor neurons (arrowheads), while SAFB and TDP-43 immunoreactivity were diffusely nuclear. b In ALS patients, motor neuron nucleolar RBM45 immunoreactivity was reduced (arrowheads) and large, skein-like cytoplasmic inclusions were observed. These inclusions were positive for TDP-43 but negative for SAFB. ALS patient glial cells frequently contained multiple RBM45 nuclear inclusions (arrows). c Glial cells from non-neurologic disease control (top) and ALS (bottom) subjects. Glial cells from ALS subjects frequently contained 1 or more nuclear RBM45 inclusions. The presence of glial RBM45 nuclear inclusions was associated with reduced SAFB immunoreactivity. d Boxplot showing the number of glial nuclear RBM45 inclusions in control and ALS subjects. For the comparison shown, * = p < 1 × 10− 8. e Stacked bar plot showing the proportion of cells (Y axis) with the given number glial nuclear RBM45 inclusions (X axis) across groups. ALS subjects have significantly more inclusions per glial cell than control subjects, who rarely exhibit RBM45 glial cell pathology. f The relationship between glial RBM45 nuclear inclusions and SAFB immunoreactivity is shown, along with the regression line. Increasing numbers of RBM45 nuclear inclusions are associated with reduced SAFB immunoreactivity, particularly at ≥3 granules per cell. The relationship between these two variables is statistically significant (p < 1 × 10−6). In the scatterplot shown, jitter is added to the points to more clearly visualize the data, but the separation between inclusion integer numbers can still be seen. The inset image shows an example of adjacent glial cells in an ALS patient where one cell without RBM45 nuclear inclusions has strong SAFB nuclear immunoreactivity, while the neighboring cell with RBM45 nuclear inclusions shows reduced SAFB immunoreactivity. In (a and b) the scale bar = 20 μm, in (c) the scale bar = 5 μm, and for the inset in (f) the scale bar = 2.5 μm

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