Fetal bovine serum gibcotm

The molecules NAT-1 and NAT-2 (Figure 1) were synthesized, purified and characterized as explained previously (Kishore et al., 2009 (link)). Fetal bovine serum (Gibco^TM) was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Casein, cellulose, methionine, Dulbecco’s Modified Eagle Medium (DMEM), penicillin–streptomycin, and trypsin–EDTA were purchased from HiMedia Laboratories (Mumbai, Maharashtra, India). Primary antibodies of AMPKα, Phospho-AMPKα (Thr172), Akt, Phospho-Akt (Ser473), p38 MAP kinase (MAPK), Phospho-p38 MAPK (Thr180/Tyr182), Phospho NF-kB were purchased from Cell Signaling Technology (Danvers, MA, USA). Cholesterol, choline bitartrate were procured from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Normal pellet diet for the animals were purchased from Amrit Feeds Ltd, Pune, Maharashtra, India. Sugar (Madhur brand) manufactured by Shree Renuka Sugars Ltd (Belgaum, Karnataka, India) was procured from the local market. The kits for biochemical estimations like triglycerides, total cholesterol, HDL cholesterol were purchased from Rapid Diagnostic Pvt. Ltd (Aspen Kits, New Delhi, India), and kits for the estimation of AST, ALT, albumin, creatinine and urea were purchased from Roche Diagnostics Ltd (Basel, Switzerland).

Primary human epidermal keratinocytes (HEKn; American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in growth medium (GM) consisting of dermal cell basal medium (ATCC) supplemented with a keratinocyte growth kit (ATCC). Cultures were maintained at 37 °C and 5% CO₂.
Human dermal fibroblasts (CCD-986Sk; ATCC) were cultured in medium composed of Iscove’s Modified Dulbecco′s Medium (Welgene, Gyeongsan, Korea), fetal bovine serum (Gibco^TM, Thermo Fisher Scientific, Rockford, IL, USA), and 1% penicillin/streptomycin (Welgene). Cells were also maintained at 37 °C and 5% CO₂.

Murine melanoma (B16-F10) was maintained in DMEM (Dulbecco’s Modified Eagle Medium) Sigma-Aldrich (São Paulo, Brazil) with 10% of fetal bovine serum (GibcoTM) Thermo Fisher (São Paulo, Brazil) and 1% streptomycin/penicillin (100 μg·mL⁻¹ and 100 UI·mL⁻¹, respectively) Sigma-Aldrich, kept at 37 °C and atmosphere with 5% CO₂.