Treatment with panobinostat enhances transcription of Nur77 or Nor1 by depleting HDAC7 occupancy at the Nur77 and Nor1 promoters. (A) HH cells were treated with the indicated concentrations of panobinostat for 4 hours. After treatment, chromatin immunoprecipitations were performed for HDAC7 and the immunoprecipitated chromatin and sonicated chromatin inputs were PCR amplified for the Nur77 and Nor1 promoters. (B) HH cells were treated with the indicated concentrations of decitabine (DAC) and panobinostat for 4 to 24 hours. Then, cells were harvested and total RNA was extracted and RT-PCR was done for Nur77 and Nor1. A β-actin–specific PCR and expression served as the loading control for the amplification. (C) HH cells were treated with the indicated concentrations of panobinostat for 4 hours. Thirty minutes before the end of the incubation, MitoTracker was added to stain mitochondria. Then, the cells were cytospun onto glass slides, fixed, permeabilized, and stained for Nur77. Images were acquired with a Zeiss LSM510 meta confocal microscope (Carl Zeiss, Heidelberg, Germany) equipped with a 63×/1.2 NA water immersion–objective lens. (D) HH cells were transfected with control siRNA or siRNA directed against HDAC7 and Nur77, respectively, for 48 hours. Following this, RT-PCR was done for HDAC7 and Nur77. A β-actin–specific PCR and expression served as the loading control for the amplification.