Increased amounts of PrPCbut unaltered mRNA levels in ADAM10 cKO mice. (A) Western blot analysis for PrPC in primary neurons of Prnp0/0, wt, ADAM10 cKO and Nestin-Cre-negative littermate controls (all from E14 embryos). Quantification of PrPC amounts shows a statistically significant increase of PrPC in ADAM10 cKO neurons compared to littermate controls (p = 0,0013). All samples were normalized to wildtype which was set to one (n = 3 for wt and Prnp0/0; n = 5 for A10 cKO and Ctrl) (d, m, u = di-, mono-, unglycosylated PrPC; representative blot is shown). Immunohistochemical staining of PrPC in brains (B) and dorsal root ganglia (C) of E14 ADAM10 cKO mice and littermate controls shows enhanced immunosignal throughout the entire central nervous system with some degree of variation between individual neurons in ADAM10 cKO mice (C, scale bar: 20 μm). Brain of an age-matched Prnp0/0 mice was taken as negative control in B. (B, upper row: whole brain section, scale bar: 500 μm; bottom row: magnification of inserts, scale bar: 50 μm). (D) Quantitative RT-PCR for PrPC mRNA in brain homogenates of ADAM10 cKO and wildtype littermate controls shows no significant difference in expression levels between genotypes at E14 (n = 4) and P1 (n = 3).