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. 2020 Oct 20;21(20):7764. doi: 10.3390/ijms21207764

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Localization of PssJ and PssJ-His₆ proteins. (A–C) Western blotting with chicken anti-PssJ IgY affinity-purified antibodies to whole-cell lysates of the wild type strain, ΔpssJ(pssJ), and ΔpssJ (A); soluble (S) and membrane fractions (M) of the wild type strain and ΔpssJ(pssJ) (B); periplasmic proteins and lysate from the spheroplasts of ΔpssJ(pssJ) (C), Western blotting with mouse anti-His6 performed to whole cell lysate (Lw), soluble (S), membrane-containing (M), membrane-associated (S_NaCl), and integral membrane (M_NaCl) protein fractions of the ΔpssJ(pssJ-his6) strain (D). Secondary antibodies conjugated with horseradish peroxidase were used, which was followed by chemiluminescent detection (A). PssO is a soluble periplasmic protein and PssP is a typical transmembrane protein with two TMS. Both served as fraction purity markers. Loading was standardized to the volume of lysate, allowing visual assessment of the cellular distribution of PssJ.