U87mg

BRAF V600E-mutant glioblastoma cell lines, AM38 and DBTRG-05MG, were purchased from the Japanese Collection of Research Bioresources (JCRB) and the American Type Culture Collection (ATCC), respectively. Control BRAF-wildtype glioma cell lines, U87MG and T98G, were purchased from ATCC. All cell lines were grown in a humidified incubator at 37 °C under 5% CO₂. NGT41, AM38, U87MG, and T98G cells were grown as adherent monolayer cultures in 10% FBS DMEM, and DBTRG-05MG cells were grown in 10% FBS RPMI1640 (Miltenyi Biotec, Aucurn, CA, USA) with Glutamax (Gibco, Paisley, UK). Dabrafenib (GSK2118436) was purchased from Selleck (Houston, TX, USA), and trametinib (GSK1120212) from AdooQ Bioscience (Irvine, CA, USA).

The human glioblastoma cell line U87/MG and human NK cell line NK92-MI were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). U87/MG cells were cultured in RPMI 1640 medium (Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and 1% penicillin–streptomycin (Hyclone). NK92-MI cells were cultured in stem cell growth medium (CellGro, Freiburg, Germany) supplemented with 2% exosome-depleted human serum (ultracentrifuged at 100,000 × g for 18 h) and 1% penicillin–streptomycin, at 37°C in 5% CO₂. U87/MG cells were transfected with a recombinant retrovirus containing a plasmid that showed enhanced expression of firefly luciferase (effluc) and thy1.1 genes, driven by a long terminal-repeat promoter (Retro–LTR–effluc–thy1.1). Thy1.1-positive cells were sorted from U87/MG cells expressing both effluc and thy1.1 genes using a magnetic cell sorter (Miltenyi Biotec, Bergisch Gladbach, Germany). Reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to confirm the expression of effluc mRNA and protein, respectively. Established stable cells expressing both effluc and thy1.1 genes were referred to as U87/MG/F cells.