Overexpression of Nur77 enhances panobinostat-induced apoptosis, whereas combined RNA interference of Nur77 and Nor1 decreases panobinostat sensitivity of CTCL cells. (A) HH cells were transfected with empty vector or pcDNA3.1 FLAG/Nur77 for 48 hours. Then, total RNA was isolated and RT-PCR was done for Nur77. A β-actin–specific PCR and expression served as the loading control for the amplification. Alternatively, total cell lysates were prepared and immunoblot analysis was done for Nur77. The levels of β-actin in the lysates served as the loading control. (B) HH cells were transfected with empty vector or pcDNA3.1 FLAG/Nur77 for 48 hours. Transfected cells were treated with the indicated concentrations of panobinostat for 48 hours. Apoptosis was assessed by staining with propidium iodide and subG1 cells were detected by flow cytometry. Columns represent mean of 3 experiments; bars, standard error of the mean. (C) HH cells were transfected with control siRNA or siNur77 and siNor1 for 48 hours. Following this, the cells were treated with the indicated concentrations of panobinostat for 4 hours and RT-PCR was done for Nur77 and Nor1. A β-actin–specific PCR and expression served as the loading control for the amplification. (D) HH cells were transfected with control siRNA or siNur77 and siNor1 for 48 hours. Following this, transfectants were treated with the indicated concentrations of panobinostat for 48 hours and apoptosis was assessed by flow cytometry. Columns represent mean of 3 experiments; bars, standard error of the mean. Statistical significance was determined by Student t test.