3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt solution

Water-soluble pharmaceutical grade of the mummy was purchased from Tuba company (http://tuba18.ir/fa/, Tehran Iran, batch number; 92-0199-s) as traditional medicine product and Figure-2 shows the product pharmaceutical packaging that illustrates the drug license from Iranian ministry of health and medical education. Roswell Park Memorial Institute (RPMI) 1640, trypsin and ethylenediaminetetraaceticacid (EDTA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) solution (Sigma-Aldrich, St. Louis, USA), phosphate-buffered saline (PBS), Fetal bovine serum (FBS), 4’,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) were obtained from Sigma Co. (St. Louis, MO, USA). Annexin V-fluorescein isothiocyanate (FITC), propidium iodide (PI) were purchased from E-bioscience (eBioscience, San Diego, CA). Cinagene Kit (RNX-Plus Solution, SinaClon, Iran), SYBR Green PCR master mix (Takara Bio Inc., Tokyo, Japan), and primers were obtained from Takapouzist Co. (Tehran, Iran). The MDA-MB-231 and MCF-7 human breast cancer cell lines and MCF-10A normal mammary epithelial cells were procured from National Cell Bank, (Pasteur Institute, Tehran, Iran).

HepG2 cells were seeded at a density of 5 × 10³ cells/well in a 96-well plate. PCBG, PCB, and MPG (purity ≥98%; Figure 1) were isolated by the chemistry department of the Chinese Material Medicine Laboratory at the Capital Medical University (Beijing, China) and QCT was purchased from National Institutes for Food and Drug Control (purity = 97.3%; Figure 1D). They were dissolved in dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA) and diluted to suitable concentrations in DMEM containing 0.5% BSA (DMSO < 0.1%). To determine the modeling concentration and the non-toxic concentration for the cells, FFA (0.6, 0.7, 0.8, and 1 mM), in addition to PCB, PCBG, and MPG (10, 50, 100, and 250 μM) were then added to each well. The plates were then incubated for 24 h at 37 °C under 5% CO₂. Then, 10 μL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution (5 mg/mL) (Sigma-Aldrich Co., St. Louis, MO, USA) was added to each well and the cells were cultured for another 4 h. The supernatant was removed and 100 μL of DMSO/well was added to dissolve the intracellular crystalline formazan product. Cell viability was determined by measuring the absorbance at 490 nm using a SpectraMax Plus 384 Microplate Reader (Molecular Devices, Sunnyvale, CA, USA).