Figure 2. Ly6C^hi inflammatory monocytes have dual features in the SILp.

(a) Comparison of microarray expression values in FACS sorted Ly6C^hi inflammatory monocytes (CD115⁺CD11b⁺), and Ly6C^hi small intestine lamina propria (SILp) monocytes (MHCII^hiCD11b⁺). Both populations were purified by FACS from T. gondii infected C57BL/6 mice at day 8 post infection.(b) Heat map representation of key pro-inflammatory and regulatory genes identified from the profiling in (a). Each column represents an individual biological replicate. (c,d,e) Intracellular cytokine staining for IL-6, IL-1α, IL-1β, TNF-α, and IL-10, in blood Ly6C^hi monocytes or SILp Ly6C^hi monocytes in whole tissue suspensions cultured for 3 hrs in the presence of brefeldin A. Cells were gated on the live Ly6C^hiCD11b^hiMHCII^hi population. (f) Enzyme immunoassay (EIA) for PGE₂ in supernatants from FACS sorted Ly6C^hi blood Mo, and Ly6C^hi SILp monocytes in whole tissue suspensions after18 hrs culture. (g) Intracellular cytokine staining for IL-10 in conjunction with iNOS, IL-6, IL-1α, IL-1β, and TNF-α in SILp Ly6C^hi monocytes cultured for 3 hr in the presence of brefeldin A. (a,b) Data are from one experiment with three biological replicates, each consisting of 1×10⁵ Ly6C^hi blood monocytes, or Ly6C^hiSILp monocytes (3–4 mice per replicate). (c-g) Data are representative of three independent experiments with three mice per group, results shown as mean ± SEM.Statistical comparisons were performed using the Student’s t test (***P<0.001).