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. 2022 Dec 21;11(1):e03298-22. doi: 10.1128/spectrum.03298-22

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Copyright © 2022 Fu et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 2 — Protein profiling of Aa isolates with serotypes a, b, or c. Different Aa isolates of serotype a, b, or c were cultured in BHI broth, and extracellular proteins were separated from the bacteria by centrifugation. Extracellular proteins were collected from the growth medium by precipitation with TCA and cellular proteins of the pelleted bacteria were extracted by bead beating. Subsequently, equal amounts of the extracellular proteins were separated by LDS-PAGE and visualized by SimplyBlue-staining (A). The presence of LtxA among the extracellular (B) and cellular proteins (C) was analyzed by Western blotting and immunodetection with a specific monoclonal antibody. LtxA-specific bands are indicated with an arrow. The serotypes of the different analyzed isolates are indicated with a, b, or c. Molecular weights of marker proteins are indicated on the left. Please note that all investigated serotype b isolates, including the isolate 30R that produces large amounts of LtxA, have the non-JP2 genotype.