Knockdown of SATB1 greatly induces IL-5 expression in polarizing Th2 cells. Cord blood CD4⁺ T cells were nucleofected with SATB1-siRNA or scrambled control siRNA and cultured in Th2-polarizing conditions. The cells and the culture media were collected at indicated time points. (A) Samples from 3 independent experiments were analyzed using quantitative RT-PCR. The normalized expression (dCt) of SATB1 mRNA of SATB1-siRNA (solid line) and control siRNA (dashed line) nucleofected cells is presented. (B) Effect of SATB1-siRNA on SATB1 expression at the 24-hour time point in the Th2 condition analyzed with Western blotting. Sc indicates scrambled control siRNA; and S1, SATB1-siRNA. Representative value of 3 independent experiments. Vertical lines have been inserted to indicate a repositioned gel lane. (C) Samples from 3 independent experiments were analyzed using quantitative RT-PCR. The normalized expression (dCt) of IL-5 mRNA of SATB1-siRNA (solid line) and control siRNA (dashed line) nucleofected cells is presented. (D) The culture media of 4 independent experiments was measured for secreted IL-5 using Bio-Plex assay and is expressed as picograms per milliliter. Each symbol type represents an independent experiment. Solid and dashed lines indicate SATB1-siRNA and scrambled control-treated cells, respectively. *P < .05. *P < .01. ***P < .005.