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. 2019 Nov 29;9:17903. doi: 10.1038/s41598-019-54494-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Lactobacilli enhanced the production of nitric oxide by macrophages. MQ-NCSU cells were seeded in 24-well plates at density 4 × 10⁵ cells/well in LM-HAHN medium and incubated at 40 °C in a humidified 5% CO₂ environment. Following incubation for 3 h, cells were stimulated with 10 or 100 MOI of a single or a mixture of heat-killed Lactobacillus spp. in DMEM and incubated at 41 °C in a humidified 5% CO₂ environment for 24 h. Supernatants were collected, and NO production was measured by Griess assay. Graphical data are presented as mean ± standard error of the mean (SEM). Bars (within a time point) which are marked by the same letter did not differ significantly (Wilcoxon signed-rank test, P > 0.05). This assay was carried out using five replicates.