Fetal bovine serum (fbs)

MDA-MB-231 was purchased from American Type Culture Collection (ATCC). MDA-MB-231-luc-D3H2LN (MDA-MB-231LN) was purchased from Caliper Life Science. The cells were maintained as a monolayer culture in DMEM, supplemented with Glutamax and penicillin-streptomycin (Invitrogen). Fetal bovine serum (FBS) (Thermo Fisher Scientific) was added to the media. Burkitt’s lymphoma cell lines, Ramos, Akata, Mutu, Raji, Jiyoye, BL-5, BL-7, and BL-8, were maintained in RPMI-1640, supplemented with Glutamax, penicillin-streptomycin, and 10% FBS (Thermo Fisher Scientific).
Whole blood was collected from healthy volunteers at James Graham Brown Cancer Center, University of Louisville with donors’ written consent. The CD4-positive and CD8-positive human T cells were purified from buffy coats via positive selection using a 1:1 mixture of CD4- and CD8- MicroBeads (Miltenyi Biotec) according to the manufacturer’s protocol. Isolated T cells were maintained in RPMI-1640, supplemented with 300 IU/mL IL-2 (R&D) and 10% FBS (Thermo Fisher Scientific). All cells were maintained at 37 °C under a humidified atmosphere of 5% CO₂.

The colon carcinoma HCT116 cell line was cultured in McCoy’s 5A (modified) medium supplemented with GlutaMAX™ (ThermoFisher), 10% fetal bovine serum (Gibco), and penicillin-streptomycin (Gibco). Breast cancer cell line MCF7 used for the gene expression analysis was cultured in Minimum Essential Eagle medium with 1 mM sodium pyruvate medium supplemented with GlutaMAX™ (ThermoFisher), 10% fetal bovine serum, and penicillin-streptomycin. HEK293 Rpn11-HTBH cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with GlutaMAX™ (ThermoFisher), 10% fetal bovine serum, and penicillin-streptomycin. B16-F10 mouse melanoma cells (ATCC) were maintained in DMEM (Lonza) supplemented with 10% FBS (Gibco) and 2 mM L-glutamine (Lonza). The cells were maintained at 37°C in a 5% CO₂ humidified incubator and were regularly passaged at 70-80% confluence. Treatments with b-AP15(Vivolux AB), bortezomib (Sigma), and chloroquine (Sigma) were all carried out at 70-80% confluence. All the compounds used were dissolved in DMSO. Treatment and control groups were exposed to an equivalent amount of DMSO (0.1-0.5%).

The human osteosarcoma cell line (U2OS), and the small cell lung cancer (SCLC) cell line (DMS114) were obtained from the American Type Culture Collection (ATCC). The non-small lung cancer cell line (HCC15) was supplied by the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures (DSMZ). U2OS cell lines stably transfected with pcDNA3.1 vector containing the sequence encoding the full-length FGFR1 (U2OSR1) or empty pcDNA3.1 vector (U2OS) were prepared as described previously (16 (link)). The U2OS cell line stably transfected with FGFR1-IIIc_K514R (U2OSR1-K514R) was kindly provided by Dr. Ellen M. Haugsten from the Department of Molecular Cell Biology, Institute for Cancer Research (Oslo University Hospital). U2OS, U2OSR1, and U2OSR1-K514R cells were cultured in DMEM (Biowest) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and antibiotics (100 U/ml penicillin, 100 μg/ml streptomycin and 1 mg/ml geneticin). DMS114 cells grew in Waymouth’s MB 752/1 medium (ATCC) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and antibiotics (100 U/mL penicillin, 100 µg/mL streptomycin). HCC15 cells were cultured in RPMI 1640 Medium (Biowest) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and antibiotics (100 U/mL penicillin, 100 µg/mL streptomycin). All cancer cell lines were kept at 37 °C in a 5% CO₂ incubator.

Human astrocyte, human glioblastoma cell lines (U87, U251, LN229, A172 and B19) and human THP-1 monocytes were obtained from ATCC (the American Type Culture Collection, Manassas, VA, USA). The human astrocyte was grown in Astrocyte Medium (AM) (ScienCell, USA) medium supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, USA). Human glioblastoma cell lines (U87, U251, LN229, A172 and B19) and murine glioblastoma GL261 cells were grown in DMEM medium supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, USA). THP-1 monocytes are maintained in culture in RPMI-1640 medium with 10% fetal bovine serum (Thermo Fisher Scientific, USA). Cell lines were tested using the ATCC cell line authentication service and routinely tested for Mycoplasma. All cells have been growing at 37°C in a humidified atmosphere (95% humidity) with 5% CO₂ (27 (link)).
In knockdown experiment, human glioblastoma cells (U87 and LN229) were treated with PLK1-siRNA (GenePharma, Shanghai, China) by using Lipofectamine RNAimax (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions. PLK1 siRNA 1#: 5’-CGAUACUACCUACGGCAAATT-3’; PLK1 siRNA 2#: 5’-CGAGGUGCUGAGCAAGAAATT-3’.

Human HT1080 cells were purchased from the American Type Culture Collection. HT1080 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific) supplemented with 10% (v/v) fetal bovine serum (FBS; Thermo Fisher Scientific), penicillin (100 U/ml), and streptomycin (100 μg/ml) (Thermo Fisher Scientific). RS4;11 cells were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ) and cultured in α-MEM with ribo- and deoxyribonucleosides (Thermo Fisher Scientific) supplemented with 10% (v/v) FBS (Thermo Fisher Scientific), penicillin (100 U/ml), and streptomycin (100 μg/ml) (Thermo Fisher Scientific). Jurkat T cells were provided by A. Rösen-Wolff and cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% (v/v) FBS (Thermo Fisher Scientific), penicillin (100 U/ml), and streptomycin (100 μg/ml) (Thermo Fisher Scientific). Murine thymocytes were isolated as described below and cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% (v/v) FBS (Thermo Fisher Scientific), penicillin (100 U/ml), and streptomycin (100 μg/ml) (Thermo Fisher Scientific). All cells were cultured in a humidified 5% CO₂ atmosphere.