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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

Fig. 6

Reduced EU-labelling in Purkinje cells of a DNA-repair deficient mouse model. a–c Confocal images (maximum projection of 10 optical sections, 4.5-μm stacks) and graphs demonstrating reduced and more variable EU labelling in Purkinje cells from 6-months-old Ercc1d/fPcp2-Cre (Ercc1) mice. Animals were perfused 1 h following EU injection. Purkinje cells (arrows) were identified with calbindin immunostaining. Bar graph in b shows the means ± SE of relative EU labelling intensities in Purkinje cells per animal (n = 3 and 4 for control and Ercc1, respectively, with at least 10 cells per mouse). Values from individual Purkinje cells are calibrated for the mean of labelling intensities of surrounding Bergman glia cells, and are shown in the violin plot in c, with the black lines indicating the median and the interquartile range (25–75% data range). The contour line represents the 95% confidence interval. Note the overall reduced nascent RNA labelling by EU and increased variation in Ercc1 Purkinje cells (c). **b P < 0.01 (unpaired Student’s t-test); ****c P < 0.0001 (Welch t-test). d–e Triple staining of EU, p53 and calbindin in Ercc1d/fPcp2-Cre cerebellum, illustrating a p53 + Purkinje cell with normal EU labelling (d), and a p53 + Purkinje cell with strongly reduced EU labelling in both the nucleolus (arrow in e, compare with cells in d) and the rest of the nucleoplasm (arrow head in e). f, g Triple staining of EU, p53 and yH2AX, illustrating a p53 + Purkinje cells with yH2AX foci (f) with low EU labelling, and a p53- Purkinje cells with yH2AX foci (g) and normal EU labelling. h–j Examples of ATF3 + Purkinje cells with relatively normal EU (h) and reduced (i, j) EU labelling. Scale bars: 20 μm

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