Dimethyl sulfoxide (dmso)

The following chemical reagents were used: gold (III) chloride trihydrate (HAuCl₄·3H₂O), trisodium citrate dihydrate (C₆H₉Na₃O₉), HS–PEG–COOH (poly(ethylene glycol), 5 kDa) from Sigma-Aldrich (St. Louis, MO, USA), and OPSS–PEG–NHS (alpha-pyridyl-2-disulfid-omega-carboxy succinimidyl ester poly(ethylene glycol), 5 kDa) from Creative PEGworks (Chapel Hill, NC, USA). Trastuzumab was isolated from Herceptin (Roche Pharmaceuticals, Basel, Switzerland). Iodogen (1,3,4,6-tetrachloro-3R,6R-diphenylglycouril) from Thermo Fischer Scientific (Waltham, MA, USA), and PD-10 column (GE Healthcare, Piscataway, NJ, USA). Hydrochloric acid and sodium hydroxide were purchased from POCH (Gliwice, Poland). Fluorescence mounting medium was obtained from Dako (Carpinteria, CA, USA). The following materials were also used in cell studies: McCoy’s, DMEM, MEM-EAGLE, and RPMI mediums and fetal calf serum from Biological Industries (Beth Haemek, Israel), phosphate-buffered saline (PBS), dimethylsulfoxide (DMSO), and the CellTiter 96^® Aqueous One Solution Reagent (MTS compound) from Promega (Mannheim, Germany). SKOV-3 and MDA-MB231 cells were also purchased from the American Type Tissue Culture Collection (ATCC, Rockville, MD, USA) and maintained based on the ATCC protocol. Additionally, all solutions were prepared using double-distilled water (18.2 MΩ·cm, Hydrolab, Straszyn, Poland).

SCC-4 cells were provided by the Ninth People’s Hospital of Shanghai Jiaotong University (Shanghai, China) and originally purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Mouse anti-human vimentin polyclonal antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Mouse anti-human E-cadherin monoclonal antibodies, rabbit anti-human Akt polyclonal antibodies, rabbit anti-phospho-Akt polyclonal antibodies and rabbit anti-human Bcl-2-interacting mediator of cell death (BIM) polyclonal antibodies were obtained from Jiamay Biotech (Beijing, China). Primer synthesis and DNA sequencing were performed by Wuhan Ying Qi Biotechnology Co., Ltd. (Wuhan, China), MTT and dimethyl sulfoxide were purchased from Promega Corporation (Madison, WI, USA), and tetramethylethylenediamine and sodium dodecyl sulfate were purchased from Sigma-Aldrich (St. Louis, MO, USA).

A172, LN18, U87MG, and U373 GBM cell lines and human embryonic kidney 293 T cells were obtained from American Tissue Type Culture Collection (USA) and cultured in DMEM medium (Gibco, USA) with %10 fetal bovine serum (Gibco, USA) and %1 penicillin-streptomycin (Gibco, USA). MGG152, GBM8-TR, and GBM8-TS cells were kindly gifted by Dr. Hiroaki Wakimoto (Massachusetts General Hospital, Boston, MA)^{19 (link)}. GBM8-TR cells were cultured in DMEM (Gibco) supplemented with 10% FBS and 1% penicillin/streptomycin. GBM8-TS cells were cultured in neurobasal medium (Gibco) supplemented with 3 mmol/L of L-Glutamine (Mediatech), B27 (Invitrogen/GIBCO), 2 μg/mL of heparin (StemCell Technologies), 20 ng/mL of human EGF (R&D Systems), and 20 ng/mL of human FGF-2 (fibroblast growth factor; PeproTech). All cells were grown in 37 °C, 5% CO₂ in a humidified incubator. TRAIL was commercially supplied (Enzo Life Sciences, US) or produced from 293T cells as described^{29 (link)}. Stock of MS-275 (Cayman Chemicals, USA) was prepared in DMSO. ZVAD-FmK and ZVA-FMK were prepared in DMSO (Promega, USA).

For the in vitro experiment, white adipocytes were treated with PA (250 nM) or bull serum albumin (BSA, 0.5%, control) for 24 h; serum-free medium or fetal bovine serum (FBS, 10%, control) medium for 12 h to induce apoptosis, and TM (1 μg/mL) for 12 h to create ERS. White adipocytes were infected with Ad-Hoxa5 or sh-Hoxa5 or 24 h or 48 h at the titer of 1×10⁹ IFU/mL. The recombinant adenovirus vector and interference vector of Hoxa5 (Ad-Hoxa5, sh-Hoxa5), adenovirus interference vector of Bax (sh-Bax) and control vectors were purchased from Gene Pharma (Shanghai, China). For signaling pathway study, the experimental procedure was as described in our previous reports in detail [43 ]. Briefly, white adipocytes were treated with Akt phosphorylation-specific inhibitor MK2206 (10 μM, Selleck, Houston, USA)and mTORC1 inhibitor rapamycin (10 μM, Selleck, Houston, USA) or DMSO (Promega, Madison, USA) for 6 h, and samples were assessed by Western blot.