Computational protocol: Repurposing Cytarabine for Treating Primary Effusion Lymphoma by Targeting Kaposi’s Sarcoma-Associated Herpesvirus Latent and Lytic Replications

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Protocol publication

[…] MM and KMM cells were seeded in 96-well plates at 5,000 cells/well for 16 h and then treated with small molecules at 5 µM final concentrations in 0.1% DMSO for 48 h. Cells treated with 0.1% DMSO and Bay11 were used as a negative control and a positive control, respectively. Cells were washed 2 times with 1× PBS and fixed with 4% paraformaldehyde for 15 min at room temperature prior to DAPI staining. Live cells were automatically counted with the Cellomics ArrayScan VTI HCS reader (Thermo Scientific), and the results were analyzed with the HCS Studio cell analysis software (Thermo Scientific). The number of live cells in the DMSO control was set as 100% and used to normalize cells treated with different compounds. A total of 73 hits that gave at least 50% cytotoxicity to KMM cells but less than 10% cytotoxicity to MM cells were selected from the first round of screening and then validated in a secondary screening, resulting in the selection of 50 final compounds. Secondary screening was carried out on the ImageXpress Micro System (Molecular Devices), and the surviving cells were automatically quantified using MetaXpress software (Molecular Devices). Compounds that showed effects similar to those of the first-round screening with less than 20% variation were selected. […]

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