Fig. 6 | Communications Biology

Fig. 6

From: Early stage prion assembly involves two subpopulations with different quaternary structures and a secondary templating pathway

Fig. 6The alternative text for this image may have been generated using AI.

Characterization of the elementary subunit of PMCA-generated PrPSc assemblies by size exclusion chromatography (SEC) under native conditions. a SEC analysis (n ≥ 3 independent fractionations) of mb-PMCA products generated with 127 S prions (10−8 dilution) immediately after the PMCA phase (day 0) or after 7 days of quiescent incubation (day 7). A representative immunoblot corresponding to elution volumes 12 ml to 18 ml is shown (electrophoretic concentration on loading as in Supplementary Fig. 1). The column calibration was performed using standard MW calibrants under identical conditions as for PMCA products analysis. b Representative sedimentogram of mb-PMCA products generated with 127 S prions (10−8 dilution) post-PMCA reaction (day 0) and after a 7-day quiescent incubation, highlighting the P1 to P2 evolution of PrPSc assemblies. c SEC profiles of mb-PMCA products generated with 127S seeds at different dilution factors, as indicated. Thirty minutes after the last sonication, the amplified products were solubilized and SEC-fractionated. The mean relative levels of PK-resistant PrPSc per fraction were obtained from the immunoblot analysis of n = 3 independent fractionations of PMCA reactions. Note the formation of at least two distinct set of assemblies as function of seed concentration

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