title: Illumina HiSeq 2500 paired end sequencing; FXR activation inhibits CAF tumor-promoting features
design: FXR activation inhibits CAF tumor-promoting features
library_name: CAFs_GW4064-treated_2_p
library_construction_protocol: CAFs at 70-80% of confluence were treated with vehicle (-) or with GW4064 (6μM) for 8-12 hrs. Then, cells were washed twice and cultured with serum-free media for the following 24 hrs. Afterwards, conditioned medium was collected, centrifuged to remove cellular debris, and used in respective co-culture experiments. Total RNA was extracted from cells using RNeasy columns (Qiagen, Germantown, MD, USA) and gene expression was assessed by real-time RT-PCR. High quality RNA from three-independent purifications for each experimental point was used for library preparation. Indexed libraries were prepared from 1μg/ea. of purified RNA with TruSeq-RNA-Sample-Prep-Kit (Illumina) following suppliers.
attributes.tag: Experimental Factor: compound
attributes.value: GW4064
attributes.tag: Experimental Factor: dose
attributes.value: 6