GSM9315382: J1; Homo sapiens; RNA-Seq
hRMCs (lots 12445, 17544, and 20018) were treated in quadruplicate wells in 6-well plates withvehicle, 25 ng/ml IL-1b, or 25 ng/ml IL-1b +20 nM eliglustat for 24, 48, or 72 hrs. Media was collected for ELISAs and GSL quantification, cell viability was measured by the alamar blue assay, and cells scraped into DPBS and replicate wells combined. The cell suspension was split into two tubes, centrifuged, and DPBS removed. Half the cells were used for GSL quantification, and the other half were used for RNA isolation. RNA was isolated using RNeasy kit with QiaShredders, resuspended in RNase free water and the concentration and quality assessed by Nanodrop. A total of 2 μg RNA per sample was shipped to Novagene for library preparation and sequencing after passing their quality control (QC) checks.