STAT5b is required for maximal production of IL-2-induced IFN-γ mRNA in TL-1 cells.

<p>STAT1 (A), STAT3 (B), or STAT5b (C) were knocked down using siRNA in TL-1 cells supplemented with IL-2. Protein lysates and RNA were harvested as well as conditioned media collected 48 h after siRNA transfection. Representative western blots of the knockdowns are shown. (D) Supernatant was analyzed to determine changes in secreted IFN-γ as a result of each knockdown. (E) The effect of knockdown on IFN-γ transcript levels was determined using qPCR. Results were normalized to the UBC gene and further normalized to the scrambled siRNA control. (F) Changes in IL-10 production were also assessed in each knockdown group using Luminex. (G) Viability following knockdown was measured by MTS assay at 48h. All results were normalized to scrambled siRNA treated cells. Student’s T test was used to determine significance compared to scrambled siRNA. * = p<0.05, ** = p<0.01, *** = p<0.005, **** = p<0.001. Data are presented as mean +/- Stdev (n = 3 biological replicates).