Cos 7 cells atcc crl 1651

COS-7 cells [ATCC CRL-1651] and NIH3T3-L1 cells [ATCC CL-173] were acquired from American Type Culture Collection (ATCC, Manassas, VA). Cell lines were validated via morphology and growth curve analysis prior to use. Cells were cultured in DMEM with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO) and 1X antibiotic/antimycotic solution (Gibco, Waltham, MA). Transfections were carried out using Lipofectamine 2000 (ThermoFisher Scientific, Waltham, MA) in 12 and 24 well plates according to manufacturer’s instructions.

The natural product, dehydroabietic acid (DA), was obtained via reported purification procedures from the commercially disproportionated rosin (Guangxi Jinxiu Songyuan Forest Products Co., Ltd.)^{53 (link)}. Its derivative which is synchronically used as the precursor in this work, DAMBA, was synthesized according to the procedures previously reported in the literature⁴¹ . All kinds of substituted salicylaldehyde (Energy Chemical, 98%), 2-bromoaniline (Energy Chemical, 98%), and chloroform-d₃ (CDCl₃, J&K Scientific, 99.8%) were used without further purification. All the organic solvents were purchased from Nanjing Chemical Reagent Co., Ltd. and used without further purifications. Milli-Q water was from a Milli-Q purification system (Merck Millipore, Germany). Dulbecco’s minimum essential medium (DMEM) and phosphate-buffered saline (PBS) were purchased from Gibco. Fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Invitrogen. Luria-Bertani (LB) broth and LB agar were from USB Co. Zinc dust. Escherichia coli (E. coli) (ATCC 25922) and Staphylococcus epidermidis (S. epidermidis) (ATCC 12228) were from ATCC. COS-7 cells (ATCC® CRL-1651™) were from ATCC