Illumina Genome Analyzer IIx sequencing; A genomewide map of PPAR_/_ target genes in human macrophages identifies a unique agonist-induced activation state - RNAseq
A genomewide map of PPAR_/_ target genes in human macrophages identifies a unique agonist-induced activation state - RNAseq
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 uM final concentration or ST247 300 nM 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. Libraries were created using the Illumina Truseq mRNA kit used according to the manufacturer's instructions.
Experimental Factor: growth condition
RPMI
Experimental Factor: compound
ST247
Experimental Factor: dose
300 nanomolar
LIBRARY_STRAND
first strand