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. 2020 Jan 15;10:3006. doi: 10.3389/fmicb.2019.03006

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Copyright © 2020 Bair and Campagnari.

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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Transwell assay to assess protection without cell to cell contact. Dual species S. pneumoniae EF3030 (Spn) and NTHi 86-028 NP (NTHi) biofilms were seeded in 24-well plates on a fixed H292 cell monolayer. To the filter inserts, approximately 10⁵ CFU per mL M. catarrhalis 7169 (solid) or CDM (open) was added. Protection was assessed at (A) 24 h and (B) 48 h where bars represent the mean and SD of three independent assays with a minimum of four technical replicates. P-value of 0.05 was considered significant and indicated by asterisk (^∗).