Cst7/cystatin F brain expression during prion pathogenesis.

A Western blotting analysis in brain extracts of terminally sick, prion-infected mice (RML), and of mice injected with non-infectious brain homogenate (NBH), showing the amount of partially protease K (PK)-resistant prion protein (PrP) (left panel). Identical protein extracts omitting proteinase K treatment were used for Western blotting with an anti-PrP and anti-actin antibody to verify equivalent loading in each lane (right panel). ZH3 denotes brain from a C57BL/6J-Prnp^ZH3/ZH3 mouse as control. Each lane denotes one mouse. B Cystatin F levels in brain extracts from the same mice depicted in A, relative to levels in NBH-injected mice (set as 100%; ***, p<0.001, Student’s t test). C Cystatin F levels in brain extracts from mice injected with either non-infectious brain homogenate (NBH) or 22L prions at different time points (dpi, days post-inoculation; term, terminal prion disease, reached at approximately 150 pdi in this experiment), relative to levels in NBH-injected mice at 150 dpi (ns, p>0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001, one-way ANOVA followed by Bonferroni’s Multiple Comparison test). D Histological analysis of cystatin F expression in cerebella of 22L-inoculated mice at the terminal stage of the disease and the relative NBH-injected controls. Three representative mice per each group are depicted. Scale bar: 100 μm.