Kd025

KD025 was purchased from Selleck Chemicals. Mitoxantrone, topotecan, cisplatin and fumitremorgin C (FTC) were purchased from Sigma-Aldrich (Merck KGaA). RPMI 1640, bovine serum albumin (BSA), fetal bovine serum (FBS), penicillin/streptomycin and 0.25% trypsin were purchased from HyClone (Cytiva). Primary monoclonal antibody against ABCG2 (cat. no. MAB4145; clone BXP-34) and AlexaFluor488-conjugated goat anti-mouse IgG secondary antibody (cat. no. A-10684) were purchased from Thermo Fisher Scientific, Inc. HRP-conjugated rabbit anti-sheep IgG secondary antibody (cat. no. AP147P) was purchased from Sigma-Aldrich (Merck KGaA). Primary antibody against GAPDH (cat. no. KC-5G4) was purchased from Aksomics Inc. [³H]-Mitoxantrone (4 Ci/mmol) was purchased from Moravek Biochemicals Inc. DMSO, MTT, DAPI and paraformaldehyde were purchased from Sigma-Aldrich (Merck KGaA). Mitoxantrone, topotecan and FTC were used in place of KD025 as positive controls to confirm the mechanism of drug resistance in leukemia cell line models. cisplatin (a non-substrate of ABCG2) was used as a negative control.

Compounds
in the DOS Informer, DOS-A, Repurposing,
and Bioactive libraries were maintained in the Broad Institute and
printed into 96- and 384-well plates using a Tecan D300e drug printer.
A subset of the repurposed compounds was purchased commercially for
validation studies: PIK-93, GSK2126458 (Omipalisib), Duvelisib (IPI-145,
INK1197), KD025 (SLx-2119), LY2784544, Palbociclib, Torin-2, AZD8186,
AT-9283, and AZD5438 (Selleckchem); ETP-45658 (R&D Systems); ETP-46464,
CP-640186, and BMS-536924 (Sigma-Aldrich); PF-03814735, TGX-221, and
Taselisib (GDC-0032) (Cayman Chemical); anandamide (VWR Scientific);
AM404 (Santa Cruz Biotech) and oleylethanolamide (Combi Blocks). Stock
solutions were prepared in DMSO and stored as per manufacturer’s
instructions.