Fig. 2: Single cell RNA sequencing (scRNAseq) and flow cytometry show increased neutrophil presence in colonic lamina propria of Lrrk2 G2019S mice following infection.
Male and female Lrrk2 G2019S and WT mice were gavaged once with ~1 × 109 CFUs of C. rodentium, and colons were harvested. Immune cells were isolated for scRNAseq and flow cytometry. A Unsupervised clustering of immune cells in WT or G2019S uninfected and infected mice. The scRNAseq datatset contains 13,892 cells total sequenced at a depth of ~20,000 genes per cell, depicted on respective UMAP plots. B Dotplot depicting identifying markers used to annotate each immune cluster based off literature. C Dotplot depicting average Lrrk2 expression across all immune cell clusters, split by condition. D Proportional analysis conducted through permutation test comparing G2019S-infected and WT-infected conditions. Significance cut off: FDR < 0.05. scRNAseq was obtained from pooled conditions of n = 3 mice per group (male-to-female ratio = 0.5). One independent experiment is presented. E Flow cytometry results of immune cell clusters projected onto t-SNE plots. F Percent Ly6G+ neutrophils isolated from CD45+ immune cells of the colonic lamina propria. Data are represented as mean ± SD and analyzed by two-way ANOVA with Fisher’s LSD post-test. n = 5–6 mice per group (male-to-female ratio = 0.9). ***p < 0.001. G MPO enzymatic assay results of colonic tissue sample measured through O.D. and normalized over gram of colonic tissue. Data are represented as mean ± SD and analyzed by two-way ANOVA with Fisher’s LSD post-test. n = 4-6 mice per group (male-to-female ratio = 0.42). *p < 0.05. One representative of two independent experiments is presented.
