Illumina HiSeq 1500 sequencing; PPARβ/δ ligand response in monocyte derived macrophages
PPARβ/δ ligand response in monocyte derived macrophages
Cells were isolated on June 06th 2013 from healthy human donor blood of unknown gender via LSM1077 gradient centrifugation. Adherent cells were positivly selected on primaria 6 well dishes (3x10^6 monocytes per well in 2 ml PBS (PAA)) followed by 6 washing steps with PBS. Finally the monocytes were cultured for 6d in humidified incubator at 37 °C, 5 % CO2 in 2ml RPMI medium supplemented with 10 % FCS (both PAA). Cells were treated with control (DMSO), L165 1 µM final concentration or PTS-264 1 µM final concentration for 1d. Cells were lysed in Peqgold (TRIzol-like solution based on phenol and chloroform). RNA was digested with DNase (MN) and purified on a Qiagen Minelute column. Illumina-compatible libraries were made using the Epicentre Scriptseq human/mouse/rat low input Gold kit.
Experimental Factor: compound
DMSO
LIBRARY_STRAND
first strand