Fig. 9

Activation of Calpain-Cathepsin in sCJD at pre-clinical stages of the disease. a qPCR analysis of expression levels of Calpain 1 and Calpain 2 in the cortex and cerebellum of the tg340-PRNP129MM mice inoculated with at sCJD MM1 at 120, 160, 180 and 210 dpi (10–1 inoculum dilution was used for animals sacrificed at 210 dpi). Fold Change represents the ratio between animals inoculated with sCJD-MM1 and control homogenates. b Western-blot and densitometry of Calpain 1, Calpain 2and Cathepsin S in the cortex and cerebellum of control (n = 3) and sCJD MM1 (n = 3) inoculated tg340-PRNP129MM mice at sCJD MM1 at 120, 160, 180 and 210 dpi (10–1 inoculum dilution was used for in animals sacrificed at 210 dpi). Fold Change represents the ratio between animals inoculated with sCJD-MM1and control homogenates. c Western-blot analysis of Calpain 1 N-ter, Cystatin C and Calpastatin in the cortex and cerebellum of control (n = 3) and sCJD MM1 (n = 3) inoculated tg340-PRNP129MM mice at pre-clinical (120dpi) and clinical (180dpi) stages. Fold change between sCJD MM1 and control inoculated animals is indicated. d qPCR analysis of expression levels of Cathepsin S in the cortex and cerebellum of the tg340-PRNP129MM mice inoculated with at sCJD MM1 at 120, 160, 180 and 210 dpi (10–1 inoculum dilution was used for animals sacrificed at 210 dpi). Fold Change represents the ratio between animals inoculated with sCJD-MM1 and control homogenates. e Western-blot analysis of PK-treated brain extracts from tg340-PRNP129MM mice inoculated with control and sCJD MM1 bran homogenates at 120 and 180 dpi. 180 dpi sample was diluted 1:5. ANOVA test followed by post-test Tukey’s Multiple Comparison Test was used to compare the values from different groups. P values for the comparisons of the three groups are indicated in the figure:*p < 0.05; **p < 0.01; ***p < 0.001