Figure 2
From: Reversible off and on switching of prion infectivity via removing and reinstalling prion sialylation
Analysis of brains and spleens from animals inoculated with SSLOW PMCAb-, dsPMCAb- or rsPMCAb-derived materials.
Syrian hamsters were inoculated IC with 10-fold diluted PMCAb-, dsPMCAb- or rsPMCAb-derived material. (A) Brain or spleen homogenates were treated with PK and analyzed by Western blot. (B) Western blots of brain homogenates from animals inoculated with PMCAb-, dsPMCAb- or rsPMCAb-derived material or non-inoculated animals 600–670 days old. Brain homogenates were treated with increasing concentrations of PK as indicated. PK-digestion profiles for three independent animals are shown. (C) Serial PMCAb reactions were seeded with 10% brain or spleen homogenates from hamsters inoculated with dsPMCAb- or PMCAb-derived material, subjected to four serial rounds and reaction products were analyzed by Western blot. As positive controls, serial PMCAb reactions were seeded with 109-fold diluted SSLOW brain material. As a negative control for cross-contamination, non-seeded serial PMCAb reactions were conducted in parallel (NS). Lane “PMCAb” refers to PMCAb reactions seeded with brain and spleen tissue from animal #8 in panel A “PMCAb-derived”. Independent serial PMCAb reactions were repeated twice for each samples and produced identical results. Black and white triangles mark diglycosylated and monoglycosylated glycoforms, respectively, whereas arrows mark the unglycosylated form. All blots were stained with 3F4 antibody.
