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. 2019 Sep 24;9:13797. doi: 10.1038/s41598-019-50310-0

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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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T-distributed stochastic neighbour embedding plots generated from canonical correlation analysis of neutrophils isolated from mouse whole blood, 24 hours after immunisation. (A) Clustering of canonical correlation analysis (CCA)-aligned neutrophils, isolated and sequenced from pooled whole blood of mice (n = 6 per group) taken 24 hours after the second dose of 4CMenB + routine and routine only immunisations. Neutrophil clusters were determined by t-distributed stochastic neighbour embedding (tSNE) dimensionality reduction during integrated analysis of CCA-aligned cells from both conditions. (B) Feature plot maps of top genes identified by differential expression analysis for each condition, with cells highlighted based on scaled expression values greater than one log₂ fold change above the median across all cells. The cells highlighted in red correspond to those expressing the top genes, Il1r2, Lcp1, and Plek, identified as positively differentially expressed in the 4CMenB + routine group relative to the routine only group and those highlighted in green correspond to those expressing the top genes, Lyz2, C1qc, C4b, identified as positively differentially expressed in the routine only group relative to the 4CMenB + routine group.