Validating lncRNA Expression Using Fluidigm's Biomark HD

We used Fluidigm’s Biomark HD to validate lncRNA expression. Pre-amplified samples and primers were loaded onto a 96 by 96 integrated fluidic circuit (IFC) chip (Fluidigm, USA). Gene expression was quantified by the relative standard curve method^{31 (link)}, and using the SsoFast EvaGreen Supermix (Bio-Rad, 1725210, USA). LncRNA expression was normalized against the geometric mean of 3 HKG’s: ARHGEF12, B-ACTIN, and TUBA1A. Each of these 3 housekeeping genes was stable across conditions in both the RNA-seq and qPCR data (Supplementary Table 3). To assess their stability in qPCR data, each housekeeping gene was normalized to the geometric mean of the other 2 housekeeping genes.