Figure 3. Up-regulation of CTSE by Ebi3^−/−Il10^−/− T_regs.

(A) mRNA was isolated from wild type or knock out T_reg purified by FACS and used for Affymetrix analysis. Modulated genes in knock out T_reg compared to wild type T_regs is depicted in a heat map. (B) Volcano plot comparing wild type and Ebi3^−/− Il10^−/−, T_regs. Highest modulated genes are marked. (C) mRNA was isolated from wild type or knock out T_reg purified by FACS, cDNA synthesized and Ctse expression assessed by qPCR. Data are the mean of 2 independent experiments. (D) Wild type or knock out T_reg were stained for intracellular CTSE [grey - 2^nd antibody control, open histograms; in green - wild type T_regs and in blue; Ebi3^−/− Il10^−/− T_regs]. (E) Equal numbers of FACS purified wild type or knock out T_reg were lysed, CTSE immunoprecipitated and analysed by SDS-PAGE/western blot. Data are representative (A, B, D and E) of three independent experiments.