Sialylation of Prion Protein Controls the Rate of Prion Amplification, the Cross-Species Barrier, the Ratio of PrPSc Glycoform and Prion Infectivity
Figure 6
Correlation between PrPSc sialylation status and its infectivity.
A. The following material was analyzed by 2D and animal bioassay: 1% brain-derived 263K material, PMCAb-derived 263K (products of 24th PMCAb rounds conducted in NBH with 10-fold dilution between rounds), PMCAb-derived 263K produced in dsNBH (products of 7th PMCAb round conducted in dsNBH with 1000-fold dilution between rounds) and 263KR+ (263K brain material subjected to 12 serial PMCAb rounds in RNA-depleted NBH and then an additional 14 PMCAb rounds in NBH as described in [46]). Prior to inoculation, all PMCAb-derived materials were diluted 10-fold. 104-fold diluted 263K brain material was used for inoculating a reference group to match the amount of PK-resistant material in PMCAb-derived samples. Diglycosylated and monoglycosylated PrPC are marked by black and white triangles, respectively. B. Western blots of 263K- or SSLOW-seeded PMCAb-derived material produced using NBH or dsNBH and treated with increasing concentrations of PK as indicated. All blots were stained with 3F4 antibody.