Illumina HiSeq 4000 sequencing; Novel RORg inverse agonists do not modulate androgen receptor activity in prostate cancer
Novel RORg inverse agonists do not modulate androgen receptor activity in prostate cancer
LNCAP cells were seeded at an appropriated density and dosed 24 hours later with 1uM of each compound. Either 24 or 48 hours later cells were lysed in RLT buffer (Qiagen) on ice; adherent cells were scraped and the material was passed through a QIAshredder (Qiagen). Samples were then snap frozen on dry ice and stored at -80ºC. Samples were randomised and extracted on Qiacube-HT following the RNeasy 96 Qiacube HT total RNA Cell with DNase protocol. RNA concentration and quality was assessed with Qubit RNA BR kit (Invitrogen) and Agilent RNA 6000 Nano kit.  Libraries were prepared using the TruSeq Stranded mRNA library preparation kit as per the manufacturers guidelines