High-Throughput Screening of Small Molecule Modulators

B16.F10 and B16.gp33 (Kindly provided by Dr. H.P. Pircher), MC-38, A549, MDA-MB-231 cell lines were maintained Dulbecco Modified Eagle's Medium (DMEM). Human RPMI-7951 cells were maintained in Eagle’s MEM. MOPC cells were cultured as previously described [70 (link)]. Media was supplemented with 10% FCS (15% for SK-MEL24) and penicillin/streptomycin. Cells were incubated at 37 °C in 5% CO2, and routinely confirmed to be mycoplasma-free (MycoAlert Mycoplasma Detection Kit, Lonza). For screening, the NIH Clinical Collection (NCC) composed of 770 small molecules was used in the screen. 5-Nonyloxytryptamine, Sumatriptan, Forskolin (all from Sigma), Asenapine and the p-CREB 3i inhibitor (SelleckChem) were dissolved in DMSO. Serotonin was dissolved in water (SelleckChem).