Dulbecco s phosphate buffered saline

Graphite flakes (Alfa Aesar, 99.8%, natural graphite 325 mesh), 6-arm PEG-amine (JenKem Technology, 15 kDa), sulfuric acid (H₂SO₄, Rankem, 95%−98%), potassium permanganate (KMnO₄, Aldrich, 99%), phosphoric acid (H₃PO₄), hydrogen peroxide (30% w/v) and potassium hydroxide (KOH, powder) from Finar Scientific, Vatalanib (cat#A3969, APExBIO), CPI444 (cat#1202402-40-1, MedChemExpress), and GBM U87 cell lines were obtained from the National Centre for Cell Sciences (NCCS) Pune. The origin of GBM U87 (or U-87 MG) is a malignant glioma from a human male patient with epithelial morphology. It is from the primary brain tumor and grade IV glioma according to the WHO classification (29 (link)). MEM (Minimum Essential Medium Eagle, Cat#AL080A) media, trypsin-EDTA solution 1× 0.25% (Cat#TCL007), FBS (Fetal Bovine Serum, Cat#RM10681), antibiotic solution (penicillin and streptomycin), and PBS (Dulbecco’s Phosphate Buffered Saline, powder pH 7.4) were purchased from HiMEDIA, India. Fluorescein isothiocyanate (FITC, Invitrogen) and CCK-8 (Cell Counting Kit-8/Cytotoxicity assay kit) were purchased from Dojindo Molecular Technologies, USA.

Experiments were performed at the National Institute of Immunology, New Delhi. 20 ml of peripheral blood was collected from five healthy volunteers aged between 25–30 years, after obtaining their written informed consent. Blood was collected more than once from some of the volunteers and there was a minimum gap of three months between two successive sample collections. The peripheral blood mononuclear cells (PBMCs) were isolated from blood by density gradient centrifugation using HiSep
^TM LSM 1077 (Himedia, Mumbai; India). The obtained PBMCs were washed thrice with Dulbecco’s phosphate buffered saline (Himedia, Mumbai; India) and counted using the trypan blue dye exclusion method with a hemocytometer (Rohem Industries Pvt Ltd, India).