Fig. 6
ES-62 modulates the gut-bone marrow axis during the early phases of CIA. a Changes in the indicated bacterial populations in the colon faecal matter of Naive, PBS- or ES-62-treated animals were measured at day 6 post CII/CFA-immunisation by qPCR and data were normalised to total bacterial content and presented as fold change compared to Naive mice (Naive, n = 3; PBS-CIA, n = 3 and ES-62, n = 2). b Representative H&E images of ileum and colon pathology of mice culled at day 6 of CIA. Scale bars are 200 µm. c IL-17+ lymphocytes were determined by flow cytometric analysis of MLN cells from individual mice (naïve, n = 5: PBS, n = 5 and ES-62, n = 5). Statistical significance was determined using one-way ANOVA with LSD Fishers multiple comparisons and significance indicated by asterisks, *p < 0.05 and ***p < 0.001. d Whole bone marrow was used to quantify MyD88 mRNA levels using qRT-PCR and normalised to naive controls. e The proportion of monocytes (CD3-B220-Ter119−Ly6G−Ly6C+) in bone marrow was measured by flow cytometry and normalised to those in Naive control mice. f, g Osteoclasts (OCs) were differentiated from bone marrow obtained at cull and cultured for 5 days and size of OCs (f) was measured using ImageJ analysis software and normalised to those from Naive controls with representative images for each disease stage in both treatment groups displayed (g). Statistics: data are presented as mean ± SEM values from individual animals (d; Naive; n = 4, PBS; n = 8, ES-62; n = 6, Naive + ABX; n = 4, PBS + ABX; n = 8, ES-62 + ABX; n = 8. e Initiation; n = 6, Pre-Clinical; n = 3, Disease; n = 4) or mean ± SD from experimental replicates (f; Initiation; n = 18, Pre-Clinical; n = 9, Disease; PBS: n = 12 and ES-62; n = 15). Statistical significance was determined using two- (e, f) or one-way (d) ANOVA with LSD Fishers multiple comparisons and indicated by asterisks, *p < 0.05, **p < 0.01 and ***p < 0.001
