Roles of butyrate and TSA on IL-2-mediated signalings in ILT-Mat cells.

(A) Annexin V-FITC staining of ILT-Mat cells. ILT-Mat cells were incubated with 200 nmol/L TSA for 18 (stippled line), 27 (dashed line), or 36 hours (solid line) and stained with annexin V-fluorescein isothiocyanate (FITC). Untreated cells were also stained (thin line). Shown is a representative experiment. (B) HDAC inhibitors did not affect IL-2–mediated elevation of tyrosine phosphorylation levels in SHP-2. ILT-Mat cells were starved for 20 hours and treated with IL-2 (+) for 10 minutes with or without preincubation of 2 mmol/L butyrate (Bu) or 500 nmol/L TSA (TSA) for 0.5 hours. (C) HDAC inhibitors did not affect IL-2-mediated tyrosine phosphorylation of Jak1 and STAT5. ILT-Mat cells deprived of IL-2 for 20 hours were treated with IL-2 (+) for 15 minutes with or without preincubation of 2 mmol/L butyrate (Bu) or 500 nmol/L TSA (TSA) for 0.5 hours. SHP-2, Jak1, or STAT5 was immunoprecipitated from the lysates of ILT-Mat cells (lower panels) and their tyrosine phosphorylation levels were analyzed by immunoblotting using 4G10 (upper panels). The positions of SHP-2, Jak1, or STAT5 are indicated by arrows.