Figure 6.

Increased renal Th1 and Th17 immune response in nephritic mice lacking IL-10–producing Tregs. Renal single cell suspensions from non-nephritic Il10^flox/flox control mice and nephritic Il10^flox/flox mice and nephritic Foxp3^YFP-Cre x Il10^flox/flox mice were stimulated in vitro with PMA/ionomycin 7 days (A and B) or 14 days (C and D) after NTN induction. Intracellular cytokine production of IFNγ and IL-17 was analyzed by flow cytometry. Representative dot plots are depicted. Cells are gated on CD4⁺ T cells, and numbers represent events in quadrants in percentage of all gated events. The frequencies of IFNγ–/IL-17–producing renal CD4⁺ T cells were quantified (*P<0.05, ***P<0.0001). (E and F) Total RNA was extracted from kidneys. Subsequently, quantitative real-time RT-PCR was performed for IFNγ, IL-17, RORγt, and CCR6 expression 7 days or 14 days after NTN induction (*P<0.05, **P<0.01, ***P<0.0001). The expression levels are indicated as x-fold of non-nephritic Il10^flox/flox control mice. Error bars represent standard error of the mean (SEM).