Penicillin streptomycin stock solution

Human osteoblast hFOB1.19 cells and human osteosarcoma U2OS and SAOS-2 cells were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in the Dulbecco's Modified Eagle's Medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and 1% penicillin/streptomycin stock solution (Beyotime, Shanghai, China). Cells were maintained in a humidified atmosphere containing 5% CO₂ at 37℃. U2OS and SAOS-2 cells (80% confluence in 6-well plates) were treated with different concentrations (0, 0.1, 0.25, 0.5, 1, 2.5, 5, or 10 µg/mL) of epirubicin (MedChemExpress, Princeton, NJ, USA) for 24 or 48 h. U2OS and SAOS-2 cells were transfected with miR-NC mimics, miR-1301 mimics, miR-NC inhibitors, miR-1301 inhibitors, pcDNA vector (vector), and TRIAP1 overexpression plasmid (TRIAP1). After 24-h transfection, cells were treated with epirubicin (1 µg/mL) for 48 h. The transfected or treated cells were used for subsequent experiments.

Poly(vinylpyrrolidone) (PVP K30), ferricyanide (K₃[Fe(CN)₆]·3H₂O), hydrogen chloride (HCl), zinc acetate (Zn(OAc)₂), trisodium citrate (Na₃(C₆H₅O₇)·2H₂O), monosodium phosphate, disodium phosphate and glutathione (GSH) were purchased from Aladdin Reagent, Ltd (Shanghai, China). Dulbecco's modified Eagle medium (DMEM) was purchased from Cytiva (USA). Fetal bovine serum (FBS) was purchased from ABW (China). Penicillin-streptomycin stock solution was purchased from Beyotime Biotechnology (China), Calcein-AM/Propidium Iodine (PI) Live/Dead Cell Double Staining Kit was purchased from Bestbio (China). Annexin V-FITC/PI Apoptosis Detection Kit was purchased from 4A Biotech (China). Anti-rabbit GAPDH antibody (10494–1-AP) was purchased from Proteintech Group (USA). Anti-rabbit mutant p53 antibody (mutp53, Y5, ab32049) and HRP conjugated anti-rabbit IgG secondary antibody (ab6721) were purchased from Abcam (USA).