Figure 2.

CX3CR1 promotes Ly6C^high monocyte adhesion and prevents renal damage during sepsis. (A) Photomicrographs and (B) quantification of kidney histologic lesions 24 hours after CLP in WT (blue) and Cx3cr1^−/− mice (red). Bars represent mean±SD (n=10 WT, n=8 Cx3cr1^−/−, from at least two repeated experiments; ANOVA with Bonferroni adjustment was used; ***P<0.001). (C) Urea and creatinine measurements in plasma of CLP-operated WT and Cx3cr1^−/− mice bars represent mean±SEM (n=10 WT sham, 6 Cx3cr1^−/− sham, 15 WT, and 15 Cx3cr1^−/− CLP, from at least two repeated experiments; ANOVA with Bonferroni adjustment was used; *P<0.05). (D) Number of Ly6C^high monocytes in bone marrow, blood, and kidney of WT (blue) and Cx3cr1^−/− (red) mice. Bars represent mean±SD (n=10 WT sham, 5 WT CLP at 6 hours, 12 WT CLP at 24 hours, and 6 Cx3cr1^−/− sham, CLP at 6 hours, and CLP at 24 hours, from at least two repeated experiments; ANOVA with Bonferroni adjustment was used; *P<0.05). (E) Two-photon laser scanning microscopic images with overlay of monocyte migratory tracks (pink) in kidney cortex of MacBlue×Cx3cr1^gfp/+ and MacBlue×Cx3cr1^gfp/gfp mice, 6 hours after CLP. ECFP signals are in cyan, GFP signals are in green, renal tubules are autofluorescent, and blood vessels are visualized by 2 MDa rhodamine-dextran. (F) Relative frequency of the three monocyte behaviors in MacBlue×Cx3cr1^gfp/+ and MacBluexC×3cr1^gfp/gfp mice. Bars represent mean±SEM (n=3 MacBlue×Cx3cr1^gfp/+; n=4 MacBluexC×3cr1^gfp/gfp from independent experiments; ANOVA with Bonferroni adjustment was used; **P<0.01). (G) Dwell time and contact duration with the renal endothelium of ECFP⁺Cx3cr1^gfp/+ and ECFP⁺Cx3cr1^gfp/gfp monocytes. Black bars indicate means. (H) Radar chart representation shows ECFP⁺ cell dynamic signatures in CLP-operated MacBlue×Cx3cr1^gfp/+ (red) and MacBlue×Cx3cr1^gfp/gfp (blue) mice. Mean values are presented within the 95% confidence interval of the measured value scale for each parameter (for all two-photon experiments, n=3 MacBlue×Cx3cr1^gfp/+; n=4 MacBluexC×3cr1^gfp/gfp from independent experiments; Mann-Whitney test for multiple comparisons was used; ***P<0.001). (See also Supplemental Figure 2 and Supplemental Movie 5).