Fig. 1

Cerebellar neurons are differentially vulnerable to bilirubin. a Appearance of a jaundiced Ugt1 ^-/- mouse (Ugt1 ^-/-, red arrowed) and a wild-type (WT) littermate at P8. b WB analysis of total cerebellum protein extract using anti-cleaved and anti-total caspase3 antibodies at P5 and P8 of WT and Ugt1 ^-/- mice. The bar graphs show the mean of cleaved/total caspase3 ratio of the bands. c WB analysis and quantification of total cerebellum protein extracts of WT and Ugt1 ^-/- mice using an anti-calbindin antibody at the indicated time points. d Representative fluorescent immunohistochemistry of cerebellar sections from WT and Ugt1 ^-/- mice at P8 and P10, using an anti-calbindin antibody (red) to highlight PCs. Hoechst (blue) was used to mark nuclei. Scale bar 50 μm. The quantification of PC number at P8 and P10 of WT and Ugt1 ^-/- mice is represented in the bar graph (cell/mm). e Left panel, Nissl staining of cerebellar layers at P10 of WT and Ugt1 ^-/- mice. Scale bar 100 μm. Right panel, layer depth quantification of P8 and P10 WT and Ugt1 ^-/- (μm). f WB analysis of total cerebellum protein extracts of WT and Ugt1 ^-/- mice at P10 using an anti-NeuN antibody. g Representative fluorescent immunohistochemistry of cerebellar sections from WT and Ugt1 ^-/- mice using anti-NeuN antibody (green) to stain differentiated granule cells. Hoechst dye was used to stain nuclei (blue). Scale bar 50 μm. For all the experiments, values represent mean ± SD. Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001. The number of WT and Ugt1 ^-/- was ≥3 in all the experiments and time points. For WB analysis, β-tubulin was used as loading control. EGL external germinal layer, IGL internal granular layer, ML molecular layer