Altered intracellular distributions of acetylated α-tubulin, mitochondria and peroxisomes. Altered intracellular distributions of acetylated α-tubulin,

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Altered intracellular distributions of acetylated α-tubulin, mitochondria and peroxisomes. Altered intracellular distributions of acetylated α-tubulin, mitochondria and peroxisomes. (A) Images of cells showing automated image analysis. Top left panel: Original image of two cells with nuclei (blue, DAPI), acetylated α-tubulin (green, immunofluorescence) and peroxisomes (yellow, PEX14-immunofluoresence). Top right: nuclei (green) identified. Bottom left: Cell boundaries identified; the non-green cell was rejected because it overlaps the edge. Bottom right: Peroxisomes (circles) identified. (B) Acetylated α-tubulin immunofluorescence (stable microtubules) within cellular subregions (outer, middle and inner). Patient cells (shaded bars) expressed significantly less acetylated α-tubulin overall than controls (open bars) (P<0.0001) and less in each region. Examples of images of acetylated α-tubulin immunofluorescence are shown in supplementary material Figs S5, S6. (C) MitoTracker fluorescence (mitochondria) within cytoplasm subregions (outer, middle and inner). Patient cells (shaded bars) had significantly less fluorescence overall than controls (open bars) (P<0.05) with the outer region more greatly affected. (D) PEX14 immunofluorescent peroxisomes within cytoplasm subregions (outer, middle and inner). Patient cells (shaded bars) had significantly more peroxisomes overall than controls (open bars) (P<0.05) with the outer region more greatly affected. Data indicate mean ± s.e.m. Post-hoc Bonferroni pair-wise comparisons: *P<0.05, **P<0.01, ***P<0.001. Greger Abrahamsen et al. Dis. Model. Mech. 2013;6:489-502 © 2013. Published by The Company of Biologists Ltd