The microarray analysis of gene expression profile. RNA was purified from pltLys-iTreg, purTGFβ-iTreg, and Tconv cells. Samples were labeled and hybridized to Affymetric 430 2.0 GeneChips. Data are averaged from 3 arrays for each subset. Image data were analyzed with Affymetrix Expression Console software and normalized with Robust Multichip Analysis to determine signal log ratios. (A) Venn diagram showing commonly and uniquely regulated probe sets found in pltLys-iTreg and purTGFβ-iTreg cells. Probe sets that revealed a 1.4-fold or greater difference (|log2 ratio| > 0.5) and ranked a product false discovery rate of <10% relative to Tconv cells are shown. (B) Heat map showing the fold change in expression of the 1646 differentially regulated probe sets identified in panel A. (C) Annotated heat map showing the expression levels of selected differentially regulated probe sets. For panels B and C, the scale (−4-fold to +4-fold). (D) Bar graphs compare the fold change in expression of select prototypical genes associated with immune functions in pltLys-iTreg and purTGFβ-iTreg cells. (E) Flow cytometry analysis of GzmB (left) and IFN-γ (right) expression in pltLys-iTreg and purTGFβ-iTreg cells.