Human u87 glioblastoma cells

Human U87 glioblastoma cells (ATCC, Manassas, VA, USA) were cultured in Eagle’s Minimal Essential Medium (EMEM; ATCC, Manassas, VA, USA) containing L-glutamine (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and 1% penicillin/streptomycin antibiotic (Thermo Fisher Scientific, Waltham, MA, USA). Human U251 glioblastoma cells (NCI Developmental Therapeutics Program, Bethesda, MD, USA) were maintained in EMEM containing L-glutamine supplemented with 10% FBS, 1% non-essential amino acids (Thermo Fisher Scientific, Waltham, MA, USA), and 1% penicillin/streptomycin.

Materials for the preparation of the hydrogels, acrylamide (AAm, 40% w/v), N,N′-methylenebisacrylamide (Bis, 2% w/v), alginic acid (sodium salt, low viscosity), ammonium persulfate (APS), N,N,N′,N′-tetramethylethylenediamine (TEMED), and calcium chloride (CaCl₂) were purchased from Sigma Aldrich (Saint Louis, MO, USA), and agarose (low melting) was purchased from Thermo Fisher Scientific (Waltham, MA, USA), and used as received. Tris-HCl buffer (pH 7.2) was purchased from Life Technologies (Carlsbad, CA, USA) and binzil silica nanoparticle colloid solution with a mean particle size of 4 nm was a gift from AkzoNobel Pulp and Performance Chemicals Inc. (Marietta, GA, USA). 5-FU was purchased from Sigma (Saint Louis, MO, USA). Human U-87 glioblastoma cells were obtained from ATCC (Manassas, VA, USA), Dulbecco’s modified Eagle medium (DMEM) from Mediatech (Manassas, VA, USA), fetal bovine serum and penicillin-streptomycin from Invitrogen (Carlsbad, CA, USA), and sodium pyruvate, MEM non-essential amino acids and GlutaMax from Life Technologies (Carlsbad, CA, USA). WST cell proliferation assay kit was purchased from Dojindo Molecular Technologies (Rockville, MD, USA).

Human U87 glioblastoma cells, obtained from the American Type Culture Collection (ATCC, Manassas, VI, USA), were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco-Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; Gibco-Thermo Fisher) at 37 °C in a humidified 95% air and 5% CO₂ atmosphere. Cells were transfected with expression plasmid pCMV6-ZNF554 (OriGene, Rockville, MD, USA) or empty vector pCMV6 (OriGene) using Lipofectamine-2000 (Thermo Fisher Scientific) following the manufacturer’s instructions. Transfection media was replaced with fresh medium six hours post-transfection. Cells were split into two batches 24 h after transfection, harvested 48 h after transfection, washed twice with ice-cold Dulbecco’s PBS, and then pellets were stored at −80 °C until use.