Figure 2.

Default of conventional DCs and monocytes-derived cells recruitment in NOD2^-/- mice under microbiota deprivation. Colon Lamina Propria Mononuclear Cells (LPMC) frequency analysis in Wild-type (WT) (blue) and NOD2^-/- (red) mice raised under Specific-Pathogen Free (SPF)(sharp color) and Germ-Free (GF)(light color) conditions. (A) Gating strategy to determine the frequency of conventional DC1 (CD11c⁺CD11b^-), of mo-DCs (CD11c⁺CD11b⁺Ly6C⁺CCR2⁺MHCII⁺), of CD11c⁺ Macs (CD11c⁺CD11b⁺Ly6C⁺CCR2^-MHCII⁺), of conventional DC2 (CD11c⁺CD11b⁺Ly6C^-CCR2^-MHCII⁺), and the frequencies of mo-Macs (CD11c^-CD11b⁺Ly6C^-CCR2^-MHCII⁺), and Mo^-MHCII⁺ (CD11c^-CD11b⁺Ly6C⁺CCR2⁺MHCII⁺), based on their CCR2, Ly6C and MHC levels, after exclusion of Lineage and doublet cells. (B) The frequency and MHCII GeoMean of CCR2⁺Ly6C⁺MHCII⁺ activated monocytes were evaluated in the monocyte-derived cells CD11c^-CD11b⁺. (C) The frequency of mo-DCs was evaluated in the Ly6C⁺ cells. The frequency of CCR2^-Ly6C^- mo-Macs was evaluated in the CD11c^-CD11b⁺ monocyte-derived cells. (D) Frequency of DC1 cells (CD11c⁺CD11b^- gate) and DC2 cells (Ly6C^- gate). Bars indicate mean ± SEM (n=4-6/group). Statistical significance was assessed by two-way ANOVA. *P<0.05; ***P<0.001.