Multiplex Gene Expression-Based Genome-Scale siRNA Screen to Identify Tumor-Specific Vulnerabilities in HCT116 Colon Cancer Cells - Geomean Normalized Data
- Deposit:2018-08-15
- Hold-Until:2019-05-25
- Modify:2018-08-15
HCT116 colon cancer cells were reverse transfected in 384-well plates with a final concentration of 50 nM siRNA in a transfection volume of 30 microliter containing 2,500 cells/well with 0.05 microliter Dharmafect 4 per well.
Gene expression for BNIP3L, NDRG1, ALDOC, LOXL2, BNIP3, and ACSL5, as well as control genes HPRT and PPIB was measured using branched DNA signal amplification via the Quantigene 2.0 assay (Affymetrix) based on the manufacturer's protocol.
The data has been background subtracted using "blank" wells (40 microliter of water and 30 microliter of hybridization solution) and geometric mean (geomean) normalized to correct for cell number based on the housekeeping genes (PPIB and HPRT). Some controls, including the blank wells, wells containing only Dharmafect 4 (DF4) reagents, and lethal controls (siRNA targeting PLK1 or PPIB), have been removed.
This PubChem submission contains the geomean-corrected (based on HPRT and PPIB reads) data for this assay. See PubChem additional PubChem submissions for the raw (lewis_rnai_screen_raw) and normalized/analyzed (lewis_rnai_screen_normalized_results) data.
PUBCHEM_ACTIVITY_OUTCOME was set to 'Unspecified' for all experimental wells and 'Inactive' for all controls including negative controls (those treated with nontargeting siRNA) and positive controls (those treated with KSR1 targeted siRNA).