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. 2018 Sep 26;9:2294. doi: 10.3389/fmicb.2018.02294

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Copyright © 2018 Kánová, Jiménez-Munguía, Majerová, Tkáčová, Bhide, Mertinková, Pulzová, Kováč and Bhide.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Validation of interaction between selected protein candidates of N. meningitidis and cell lysate of hBMECs by semi-quantitative ELISA and immunocytochemistry. (A) Semi-quantitative ELISA performed to confirm the interaction between proteins of hBMECs and bacterial ligands (different concentrations). The interaction was detected with HisProbe conjugated with HRP and is shown in Relative Fluorescence Units (RFU). As positive control (+ control)–domain III protein E of West Nile virus was used. Negative control: (– control)–no ligand was added. Input control: recombinant ligand NMB0375 was coated and incubated with HisProbe-HRP and substrate. (B) Interaction of selected proteins of N. meningitidis with cultured hBMECs. Nuclei are stained with DAPI. Negative control–no recombinant ligand was included in the assay. Scale bars−10 μm. In both panels: NMB0375-adhesin MafA1; NMB2039-major outer membrane protein P.IB; NMB1994-putative adhesin/invasion; NMB1126-putative lipoprotein; NMB1946-outer membrane lipoprotein. Assays were performed in triplicate.