Mm 1s

The following cell lines (ATCC, except where noted) were grown in culture medium supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco), 100 units/mL penicillin, 100 μg/mL streptomycin (Gibco), and 1x GlutaMAX (Gibco) (except where noted), and maintained in a humidified incubator at 37°C in 5% CO₂; Rev-GFP U2OS [McCoy's 5A, 200 ug/ml G418 (Sigma)], MM.1S (RPMI), MV-4–11 (IMDM), THP-1 (RPMI), HCT-116 (McCoy's 5A), AML2 (DSMZ, RPMI), AML3 (DSMZ, RPMI), HT1080 (EMEM), HEL (DSMZ, RPMI), Kasumi-6 (RPMI, 2 mM L-glutamine, 1.5g/L sodium biocarbonate, 4.5 g/L glucose, 10 mM HEPES, 1.0 mM sodium pyruvate, 2 ng/mL GM-CSF, 20% FBS), SINE compound resistant HT1080 (EMEM, 600 nM KPT-185), A549 (RPMI), UCH1 (4:1 IMDM:RPMI), UCH2 (4:1 IMDM:RPMI), LS174T (EMEM), and ASPS-KY (gifted from A. Ogose, RPMI). The XPO1 SINE™ compounds KPT-330 (selinexor), KPT-335, KPT-350, KPT-8602, KPT-301, KPT-9159 (biotinylated LMB, b-LMB), KPT-9058 (biotinylated KPT-276), and KPT-9511 (biotinylated KPT-8602) were synthesized at Karyopharm Therapeutics, Inc. (Newton, MA). LMB was purchased from Cell Signaling.

Jeko-1, Mino, MM.1S, and HEK293T cells were bought from the American Type Culture Collection (ATCC), and DAOY cells were a gift from Dr. Alex Huang (Case Western Reserve University, OH, USA). Jeko-1, Mino, MM.1S, and DAOY cells were cultured in RPMI-1640 (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich) and 1% Pen/Strep (Hyclone). HEK293T cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (Sigma-Aldrich) with the same 10% FBS and 1% Pen/Strep supplementation. NK92 cells were a kind gift from Dr. Daniel Popkin (Innova Dermatology, TN, USA). They were cultured in Alpha-MEM media (Sigma-Aldrich) and supplemented with 12.5% FBS, 12.5% horse serum (Sigma-Aldrich), 0.02 mM folic acid (Acros), 0.1 mM 2-mercaptoethanol (Gibco), 1.5 g/L sodium bicarbonate (Fisher Scientific), 2 mM L-glutamine (Gibco), and 0.2 mM inositol (Acros). IL-2 (200 IU) was added fresh ever 2–3 days with media changes for NK92 cells. All cells were maintained at 37°C with 5% CO₂ in the cell incubator. Glucose concentrations of media used for cell culture: 2 g/L for primary NK cells, as they were in RPMI-1640 media; 1g/L for NK92 cells, as they were cultured in Alpha-MEM; and 4.5 g/L for HEK293T cells, as we use DMEM-High Glucose. All media are from Sigma-Aldrich as mentioned above.