Figure 6. CD103⁺ CD11b⁺ DCs but not CD103⁻ CD11b⁺ DCs Express IL-23p19 and IL-12p40 in Response to TLR5 Stimulation.

(A) Small intestinal lamina propria cells were isolated from six mice and pooled for FACS sorting. DAPI⁻ CD45⁺ CD11c⁺ MHCII⁺ lamina propria cells were FACS-sorted into CD103⁺ CD11b⁺ or CD103⁻ CD11b⁺ groups and individually assessed for purity by FACS analysis.

(B) TLR5 mRNA expression by quantitative PCR.

(C – D) Fold induction of messenger RNA (mRNA) encoding for IL-23 protein subunits IL-12p40 (C) and IL-23p19 (D) after 2 hours of in vitro incubation with flagellin. Data is representative of 1 out of 4 independent experiments. Error bars denote ± s.e.m.

(E) IL-12p40-YFP reporter mice received 1 μg of flagellin intravenously. Three hours after flagellin injection, mice were sacrificed and lamina propria cells were harvested for FACS analysis. Dot plots show cells gated by CD45⁺ CD11c⁺ MHCII⁺. DC subsets were analyzed for YFP expression in the following three groups: CD103⁺ CD11b⁻, CD103⁺ CD11b⁺, and CD103⁻ CD11b⁺ LPDCs. Numbers are percentage of gated cells out of the parent population (n = 3 mice per group).