Isolation and Synthesis of Mandelalides

The recollection and isolation of natural product mandelalides from Algoa Bay, South Africa and total syntheses of mandelalides A and L have been described previously [4 (link),9 (link),60 (link)]. NMR data for mandelalides tested here are included in [4 (link)] (natural products) and 61 (synthetic products), and were acquired at the start of this study; integrity of all mandelalide compounds was also routinely confirmed by LC-MS. Oligomycin A was purchased from Santa Cruz Biotech (Dallas, TX, USA). Dorsomorphin was purchased from Abcam (Cambridge, UK). Paclitaxel and salinosporamide A (marizomib) were purchased from Sigma Aldrich (St. Louis, MO, USA). Erlotinib was purchased from Enzo Life Sciences (Farmingdale, NY, USA). All chemicals were reconstituted in 100% DMSO, aliquoted and stored in amber borosilicate glass vials at −20 °C for use in biological studies. Cell culture-grade DMSO was used as the vehicle for all treatments, and final concentrations of DMSO for in vitro experiments never exceeded 0.1%. Primary and secondary antibodies were from Cell Signaling Technology, Inc. (Danvers, MA, USA). Specific codes were as follows: AMPK (#5831), pAMPKα T172 (#50081), ACC (#3676), pACC S79 (#11818) and GAPDH (#5174). General laboratory reagents were from VWR International (Radnor, PA, USA).