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Figure 2. The gut microbiome affects immune cell populations in the blood, MLN, and spleen. Circulating immune cell populations in CONV and GF mice were measured by CBC. (A) The absolute concentration and (B) the proportion of total white blood cells of circulating immune cell populations in CONV and GF mice. Flow cytometry was used to characterize immune cell populations in the (C) MLN and (D) spleen of CONV and GF mice. Representative flow cytometry plots and the gating strategy can be found in Supplementary Figure 2. PCA of the (E) concentrations and (F) proportions of circulating immune cell populations and the proportions of immune cell populations in the (G) MLN and (H) spleen in GF and CONV mice. For (A), (B), (C), and (D), statistical significance was determined using unpaired, two-tailed t-tests. Symbols represent individual samples, and lines represent the mean ± standard deviation. For (E), (F), (G), and (H), ellipses represent the 95% confidence interval, each point represents one sample, and statistical significance was determined using PERMANOVA. Data are representative of two independent experiments. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001. See also Supplementary Tables 1 and 4.