Hepg2 atcc hb 8065

Hep-3B (ATCC HB-8064) and HepG2 (ATCC HB-8065) cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, United States). Anti-gankyrin mAb (ab182576), anti-C/EBPα mAb (ab40764), and anti-PPARα mAb (ab8934) were obtained from Abcam (Abcam, Cambridge, MA, United States). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) was obtained from Santa Cruz (Santa Cruz Biotechnology, Santa Cruz, CA, United States). C/EBPα proteins were obtained from ORIGENE (Rockville, MD, United States), and PPARα was obtained from Creative Biomart (Shirley, NY, United States).

Human hepatocellular carcinoma cells (HEP-G2; ATCC HB-8065 ™), murine lymphoma cells (L5178Y-R; ATCC CRL-1722 ™), non-tumoral monkey kidney epithelial cells (VERO; ATCC CCL-81 ™), and murine macrophage (J774A.1; ATCC TIB-67 ™) cells were obtained from the American Type Culture Collection (ATCC^®, Manassas, VA, USA). Human peripheral blood mononuclear cells (PBMC) and human red cells (erythrocytes) were kindly provided by the Facultad de Medicina of the Universidad Autónoma de Nuevo León (UANL).

HepG2 (ATCC HB-8065) and AML12 (ATCC CRL-2254) cells were purchased from American Type Culture Collection (ATCC). HepG2 2.2.15 cells were kindly provided by Dr. M.-S. Ho (Academia Sinica, Taiwan). The cells were maintained in Dulbecco’s Modified Eagle Medium containing 4.5 g/l d-glucose and l-glutamine (Gibco, Invitrogen, USA) supplemented with 10% fetal bovine serum containing 1.5 g/l sodium bicarbonate, 0.1 mM nonessential amino acids, 1.0 mM sodium pyruvate, and 100 units/ml penicillin G and 100 μg/ml streptomycin. Cells were incubated at 37 °C in a humidified atmosphere containing 5% CO₂. G418 was added to the medium for maintenance of HepG2 2.2.15 cells at a final concentration of 200 μg/ml. Cells grown in various culture dishes at 80% confluence were used in all experiments.

AFB₁ and AFM₁ were procured from LKT laboratories, Inc (St. Paul MN, USA). Human hepatoma cells (HepG2; ATCC HB-8065), Eagel’s minimum Essential Medium (EMEM; ATCC 30-2003), and fetal bovine serum (FBS; ATCC 30-2020) was purchased from American Type Culture Collection (ATCC, Manassas, VA).

Hepatocellular carcinoma cells HepG2 [HEPG2] (ATCC HB-8065) and human lung carcinoma epithelial cell line A549 (ATCC CCL-185) were purchased from American Type Culture Collection (ATCC, Manassas, VI, USA). Both cell lines were cultured in DMEM without phenol red (Gibco, Milan, Italy), supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin, 100 µg/mL streptomycin (Gibco) and 2 mM L-Glutamine (Gibco). Cells were grown in an incubator at 37 °C, 5% CO₂ and 90% humidity.
Mancozeb (MZ, CAS no. 8018-01-7, purity 99%) and zoxamide (ZOX, CAS no. 156052-68-5, purity 99%) were purchased from Sigma-Aldrich (Milan, Italy), and dissolved in sterile DMSO (Sigma-Aldrich) to obtain stock solutions of 36.9 mM and 231.7 mM, respectively.
Just before use, serial dilutions of chemicals in culture medium were prepared considering the concentration used in the field by agricultural workers as 1X (starting point). For MZ, due to its limited solubility, the maximum dilution required to obtain an acceptable DMSO concentration (0.8%) corresponded to 1/10 of the field concentration (0.1X).

All chemicals were obtained from reputable suppliers: Sigma Aldrich (Saint Louis, MO, USA) provided chitosan powder [low molecular weight (LMW), deacetylation 75–85%], Merck supplied sodium tripolyphosphate (TPP), Hamburg Industries Inc. (Germany) supplied acetic acid (99.8%), and Laboratory & Scientific Enterprise (Malaysia) supplied dimethyl sulfoxide (99%). The Tween 80 and SF medications were purchased from Sigma-Aldrich. The entire experiment was conducted with deionized water (18.20 MΩ cm⁻¹). Primary Dermal Fibroblast: Normal, Human, Adult (HDFa) (ATCC® PCS-201-012™), Hepatocellular carcinoma cell, (HepG2-ATCC® HB-8065™), were purchased from ATCC (AMERICAN TYPE CULTURE COLLECTION, PO BOX 1549, Manassas, VA 20,108, USA).

Unless otherwise indicated, all chemicals and reagents were from Sigma-Aldrich, USA. Antibodies against β-actin (C4), GAPDH (6C5), Glut4 (IF8), Myogenin (5FD), and Alexa fluor 488 conjugated c-Myc (9E10) were from Santa Cruz Biotechnology, Santa Cruz, USA. Anti-Akt (pan) (C67E7), anti-phospho-Akt (Ser473) (D9E), anti-phospho-Akt (Thr308) (C31E5E), anti-IRS-1 (D23G12), anti-phospho-IRS-1 (Ser1101), anti-phospho-IRS-1 (Ser307), anti-phospho-IRS-1 (Ser318) (D51C3), anti-phospho-IRS-1 (Ser612) (C15H5), anti-IRS-2 antibody, anti-phospho-PDK1 (Ser241) (C49H2), HRP-linked anti-mouse IgG, and HRP-linked anti-rabbit IgG antibodies were obtained from Cell Signalling Technology, USA. Clarity Max™ Western ECL blotting substrates (1705062) were purchased from Bio-Rad Laboratories, Italy. The cell line C2C12 (ATCC CRL-1772), 3T3-L1 (ATCC CL-173), and HEPG2 (ATCC HB-8065) were acquired from ATCC (VA, USA).