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Supplementary Figures: Development of an in vitro assay for the detection of the polymerization of the pyrin domain of ASC

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posted on 11.06.2021, 10:28 by Ian Philipp Bresch, Dominik Alexander Machtens, Thomas Frank Reubold, Susanne Eschenburg

Supplementary Figure 1: TEV-Cleavage of MBP-ASCPYD. (A) Proteolysis of 10 µM MBP-ASCPYD with 2 µM TEV protease in buffers with 150 mM KCl and different pH values. (B) Proteolysis of 30 µM MBP-ASCPYD with 2 µM TEV protease in buffers with different pHs and different KCl concentrations. (C) Proteolysis of 200 µM MBP-ASCPYD with 2 µM TEV protease under standard conditions. The samples for B and C were diluted before applying to the gel to avoid overloading.

Supplementary Figure 2: Detection of the polymerization of ASCPYD by MST at different time points: (A): ASCPYD polymerization in solution with 600 mM KCl and a pH of 8.0 after an incubation time of 30 or 90 minutes at 37°C. (B): ASCPYD polymerization in solution with 150 mM KCl and a pH of 5.0 after an incubation time of 60 or 90 minutes at 37°C. Each experiment was performed twice.

Funding

The project was financially supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2155 “RESIST” – Project ID 39087428. IP Bresch is a graduate student in the MD/PhD program Molecular Medicine of the Hannover Biomedical Research School (HBRS).

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