Fig 2: Average kinetic curves (a) and tlag values (b) of PrP(89-230) self-replication reaction performed with introduction of PrP(107-143) seed at different time-points of the reaction. Then, 10 µL of 0.5 mg/mL PrP(107-143) fibrils were added to 100 µL of PrP(89-230) solution (0.5 mg/mL of monomeric PrP(89-230) and 0.001 mg/mL PrP(89-230) fibrils) at a different time-points. The reaction kinetics were monitored by measuring ThT fluorescence intensity (ex. 440 nm; em. 480 nm) at 60 °C through a bottom of non-biding 96-well plate (Corning No. 3881, New York, United States) using ClarioStar Plus (BMG Labtech, Ortenberg, Germany) microplate reader. The measurements were performed every three minutes. The error bars are standard deviations estimated from 3 repeats.

Fig 3: In vivo expression of PcnfY:luxCDABE fusions.(A) Schematic representation of a transcriptional (PJNS02, [20]) and a translational fusion (pBO6500) of the cnfY promoter region (PcnfY) to the luxCDABE reporter gene operon. (B) Y. pseudotuberculosis YPIII cultures harboring these plasmids were grown at 25°C and 37°C. Samples for bioluminescence measurement (described in Material and Methods) were taken after 4 hours. The data are depicted as relative light units (RLU/OD600) from three independent cultures measured on a CLARIOstar plus reader (BMG Labtech, Ortenberg, Germany). (C) Y. pseudotuberculosis YPIII cultures harboring the translational fusion including the wildtype (pBO65000), the R1 (pBO6501), or the R1+2 (pBO6502) RNAT variant were grown at 25°C and 37°C. Samples for bioluminescence measurement (described in Material and Methods) were taken after 4 hours. The data are depicted as relative light units (RLU/OD600) from three independent cultures measured on a Varioskan Flash (Thermo Scientific, St. Leon-Rot, Germany). Results from bioluminescence measurements of variant R1+2 are additionally shown as inlet on a different scale to reflect the low signal intensity.