F 12 dmem f12

Dulbecco’s modified Eagle medium/nutrient mixture F-12 (DMEM/F12), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylterazolium bromide (MTT), and 6-OHDA were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was purchased from HyClone Laboratories (Logan, UT, USA). Penicillin and streptomycin were purchased from Wako (Tokyo, Japan).
An apoptosis/necrosis detection kit (blue, green, red) and a DCFDA/H2DCFDA Cellular ROS Assay Kit were purchased from Abcam (Cambridge, UK). Antibodies such as Beclin, caspase-3, pERK, extracellular signal-regulated kinase (ERK), and LC3A/B XP, along with LumiGLO reagent, were purchased from Cell Signaling Technology (Danvers, MA, USA). RNAiso Plus was purchased from Takara Bio (Shiga, Japan). The THUNDERBIRD SYBR qPCR Mix was purchased from Toyobo (Tokyo, Japan).

The Saos-2 (ATCC HTB-85) OS cell line was provided by the bank of cell lines from human and animal tissue of the R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of the National Academy of Sciences of Ukraine. This cell line is commonly used in cancer research as a model of primary bone tumors characterized by osteoblastic features, alkaline phosphatase production, and DOX resistance [79 (link),80 (link),81 (link)]. The Saos-2 cells were cultured in Dulbecco’s modified eagle medium as follows: the nutrient mixture F12 (DMEM/F12) (Sigma-Aldrich, Taufkirchen, Germany) was supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Germany). The cells were grown with an antibiotic-antimycotic (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C and 5% CO₂ [82 ].