Figure 2.

The gut microbiota increase mature KC frequency and direct KC distribution. A: Percentage of NPCs isolated from livers of adolescent CL and GF mice expressing F4/80 (macrophage) or F4/80 and MHCII (mature macrophage) were similar. Representative of three independent experiments (n = 6 in the CL group; n = 6 in the GF group) (F4/80⁺P = 0.40; F4/80⁺ MHCII⁺P = 0.06). B: Correlative analysis revealed that mature KC frequency (F4/80⁺MHCII⁺ NPCs) was directly proportional to bacterial load (nanograms of bacterial DNA per gram of cecal stool); Pearson’s correlation coefficient = 0.73. C: Percentage of F4/80⁺ NPCs (cyan filled gate) or F4/80⁺MHCII⁺ NPCs (black lined gate) isolated from livers of adult CL mice was significantly greater than that of GF mice (n = 5 in the CL group; n = 5 in the GF group). Dot plots representative of 4 independent experiments. D: Percentage of F4/80⁺ NPCs (cyan filled gate) or F4/80⁺MHCII⁺ NPCs (black lined gate) isolated from livers of adult CL mice was significantly greater than AVMN mice (n = 5 in the CL group; n = 5 in the GF). Representative of two independent experiments. E: KC distribution in livers of CL, GF, and AVMN mice assessed by immunofluorescence staining of liver sections with F4/80 (green) and DAPI nuclear counterstain (blue). Analysis was performed by tissue cytometry (NearCYTE) (Supplemental Figure S3). Three lobular portal areas and central areas were each randomly selected from a total of three animals per group and F4/80⁺ cell numbers/0.1 mm² counted. Representative images was shown in the figure. Scale bar = 100 μm. Adult CL, GF, and AVMN mice were used. *P < 0.05, **P < 0.025, ***P < 0.01.