Rpmi 1640 media

Chemicals, solvents, pyridoxal-5′-phosphate (PLP), D,L-dithiothreitol (DTT), alliin, dialk(en)yldisulfides, amphotericin B (AmpB), fluconazole (FLC), 5-flucytosine (5-FC), kanamycin, YPD media and Sabouraud dextrose agar were purchased from Sigma-Aldrich (St. Louis, MO, USA); DEAE-sepharose was from “GE Healthcare” (Chicago, IL, USA); RPMI 1640 media and 3-(N-morpholino)propanesulfonic acid (MOPS) were purchased from PanEco (Moscow, Russia); S-alkyl-L-cysteine sulfoxides (methiin, ethiin and propiin) were synthesized according to the previously reported procedures [13 (link)]. 2-Nitro-5-thiobenzoate was obtained as described [30 ]. E. coli strain BL21 (DE3) F-ompT hsdSB gal dcm (DE3) was obtained from Novagen (Darmstadt, Germany). The plasmid with the gene of the C. freundii C115H MGL mutant form was obtained previously [13 (link)].

Human leukemia K562 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were grown in RPMI 1640 media (PanEco, Moscow, Russia) supplemented with heat-inactivated fetal calf serum (HyClone, Logan, UT, USA), 2 mM glutamine, 250 u/mL penicillin, and 250 μg/mL streptomycin (PanEco, Russia) at a temperature of 37 °C in a humidified atmosphere containing 5% CO₂. Hemin (50 μM, neoFroxx, Einhausen, Germany) was added to the medium to induce erythroid differentiation, as previously described [38 (link)], and cells were incubated further for 108 h. At this concentration, the drug did not affect cell proliferation.