Fig. 2

Analysis of relative EU-intensities in different cerebellar cell types. a EU labelling intensity depends on the distance from the EU injection site. Upper image is a widefield tile scan. Lower images are color-coded high magnification confocal images of highlighted areas, illustrating lower EU intensities in the Purkinje cell layer in lateral (a′) versus medial (a″) cerebellar cortex. b EU labelling intensities of individual Purkinje cells plotted against the mean intensities of surrounding Bergman glia. Values show a linear relationship between these values over a substantial interval of labelling intensities. Data points and linear regression lines are from four animals (3 coronal sections per animal, > 3 regions per section). Linear regression parameters in the grey box are based on all values. c Violin plot showing relative EU labelling intensities (= labelling intensity divided by mean labelling intensity of nearby Bergman glia cells) of different cerebellar cell types. d, e Confocal images (maximal projection of 6 images, 3-μm stacks), color-coded representation of EU-signal (d′), and violin plots illustrating increased relative EU labelling intensity in Purkinje cells 6 and 24 h post-injection. Nuclei of Bergman glia cells are marked by S100β (arrows in d and d′). Each dot in e represents the labelling intensity in a Purkinje cell divided by the mean of labelling intensity of surrounding Bergman glia cells. f Violin plots illustrating EU signal in granule cells relative to Bergman glia cell 1 and 6 h post injection, illustrating that signal in granule cells is absent after 6 h. Horizontal lines indicate the median and the interquartile range (25–75% data range). ****c, e P < 0.0001. (Kruskal–Wallis test with Dunn’s multiple comparison tests); ****f P < 0.0001 (Welch’s t test). Scale bars: 500 μm (a); 20 μm (a′); 10 μm (d′)