Figure 3. Th17 Cell Differentiation during Ontogeny.

Lamina propria lymphocytes were isolated from the small intestines of Rorγt gfp/+ mice at the indicated ages. For day 4 and day 16 studies, small intestines from 3-4 pups from the same litter were combined in order to obtain sufficient cell numbers for analysis. For day 25 and 33, plots represent individual mice. Top panels — surface staining and RORγt-GFP analysis; bottom panels — intracellular staining for IL-17 after 4 hour stimulation with PMA/Ionomycin in the presence of GolgiPlug (BD). Data are representative of two independent experiments with 2-3 samples/mice in each. Plots are gated on lymphocytes.