Lincode smartpool sirna library human nr lncrna refseq v54

Dharmacon Lincode^® SMARTpool^® siRNA Library-Human NR LncRNA RefSeq v54 (#G-301000) (GE Dharmacon) was used. 1×10⁴ TEAD Reporter-MCF-7 cells were plated in 96-well plates 24 hours prior to transfection. The cells were transfected with either the 15 pmol Lincode^® siRNA library or siRNA control (GE Healthcare Dharmacon). siGLO green transfection indicator was included to show that over 80% of the cells were transfected using this method. Cells were lysed 72 hours after transfection and assayed for luciferase activity using the Dual-Glo Luciferase reporter system (Promega) and activities were monitored by Synergy H4 Hybrid Multi-Mode Microplate Reader (BioTek). The protein concentrations of diluted cell lysates (1:100) were measured in parallel using Bio-Rad protein assay kit (Bio-rad), which was used to normalize the luciferase activity. The firefly luciferase activity for each 96-well plate was subtracted with background, normalized by protein concentration, and further normalized by control siRNA as fold changes. Log₂ of relative fold changes were plotted. The hits with log₂ ≤ -3 were selected as top candidates.