The human MCL cell lines Granta-519, Jeko-1, Maver-1, Mino, and Z138 were obtained from DSMZ (Braunschweig, Germany) and cultured in Iscove’s modified Dulbecco’s medium (IMDM; Invitrogen Life Technologies, Carlsbad, CA, USA). The stromal cell line HS-27a and the kidney cell line HEK-293T/17 were obtained from ATCC (Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Invitrogen).
For co-culture assays, bone-marrow stromal cells were seeded in 96-well plates 4 hours (h) prior to the addition of MCL cells, to allow cell attachment. MCL cells were pre-incubated for 2 h with stromal cells, followed by a 3-day co-culture in the presence of the indicated drug concentrations. Cell viability and specific cell death were measured by flow cytometry.
The following small-molecule inhibitors were used: venetoclax/ABT-199, Q-VD-OPh, IPTG (MedChemExpress, Princeton, NJ, USA), silmitasertib/CX-4945, S63845, MK2206 (Selleckchem, Houston, TX, USA), cycloheximide, puromycin (Sigma-Aldrich, St. Louis, MO, USA), and blasticidin (Thermo Fisher Scientific, Waltham, MA, USA).
For co-culture assays, bone-marrow stromal cells were seeded in 96-well plates 4 hours (h) prior to the addition of MCL cells, to allow cell attachment. MCL cells were pre-incubated for 2 h with stromal cells, followed by a 3-day co-culture in the presence of the indicated drug concentrations. Cell viability and specific cell death were measured by flow cytometry.
The following small-molecule inhibitors were used: venetoclax/ABT-199, Q-VD-OPh, IPTG (MedChemExpress, Princeton, NJ, USA), silmitasertib/CX-4945, S63845, MK2206 (Selleckchem, Houston, TX, USA), cycloheximide, puromycin (Sigma-Aldrich, St. Louis, MO, USA), and blasticidin (Thermo Fisher Scientific, Waltham, MA, USA).