GATA-3–dependent cytokine production in TCLs. (A) GATA-3 expression was determined by (upper) qRT-PCR and (lower) western blot in TCL lines lentivirally transduced with a nonspecific (“scramble”) or GATA-3–specific (“19301”) shRNA. (B) IL-10, (C) IL-4, (D) IL-13, and (E) IL-5 production following GATA-3 knockdown was examined by (upper) qRT-PCR or (lower) enzyme-linked immunosorbent assay. For determination of cytokine production, cells were cultured at the same density in triplicate in 96-well plates in media alone or stimulated with phorbol 12-myristate 13-acetate (50 ng/mL) and ionomycin (iono, 500 ng/mL), and cell free supernatants were collected after a 12- to 24-hour incubation. (F) The absolute eosinophil count at diagnosis is shown in PTCL, NOS patients stratified by GATA-3 expression. (G) Unsupervised clustering of PTCL, NOS (n = 31), CTCL (n = 11), and reactive lymph nodes (RLNs; n = 7) on a panel of Th1-associated (T-bet, IFN-γ, CXCR3) and Th2-associated (GATA-3, IL-4, IL-5, IL-13) transcripts. *P < .05 and P < .01 in unpaired 2-sided Student t test.