Cross-seeding of prions by aggregated α-synuclein leads to transmissible spongiform encephalopathy
Fig 2
Bioassay of dgPMCAb-derived PrPres in Syrian hamsters.
a Western blot analysis of brain material from hamsters inoculated IC with (i) fibrillar α-synuclein, (ii, iii) dgPMCAb-derived PrPres induced by WT α-synuclein fibrils or lysates of HeLa cells expressing the A30P variant, (iv) non-fibrillar α-synuclein, or (v) non-seeded dgPMCAb-derived material and stained with SAF-84 (top panels) or 3F4 antibody (bottom panels). Non-inoculated, age-matched animals were examined as a negative control. Brain materials marked by asterisks were used for the second passage. b Left panel shows Western blot analysis of dgPMCAb-derived PrPres used for inoculation and resulting brain-derived PrPres and PrPSc from animals of the 1st and 2nd passages. To match the amounts of dgPMCAb- and brain-derived PrPres on a blot, all brain materials were diluted 250-fold after PK treatment. Western blots were stained with SAF-84 antibody. On the right panel is a schematic representation of the PK resistant profile showing overlap between the three glycoforms of PrPres (gray boxes) and the three glycoforms of PrPSc (black boxes). c Western blot analysis of brain material from hamsters inoculated with the second passage of dgPMCAb-derived PrPres induced by α-synuclein fibrils; lysates of HeLa cells expressing the A30P mutant; or non-seeded dgPMCAb-derived material. Lanes marked by asterisks show brain material from animals from the first passages used for serial transmission. Western blots were stained with SAF-84 (top panels) or 3F4 antibody (bottom panels).