Synthesis and Characterization of SELENOT-derived Peptides

The SELENOT-derived peptide 43–52 (corresponding to the sequence H-Phe-Gln-Ile-Cys-Val-Ser-Sec-Gly-Tyr-Arg-OH) designated as PSELT and its inactive form (Ser 46,49, designated as the inert-PSELT or I-PSELT) were chemically synthesized as described previously [17 (link)]. Dulbecco’s Modified Eagle Medium F-12 (DMEM/F-12), Dulbecco’s phosphate-buffered saline (DPBS), penicillin/streptomycin, 0.25% Trypsin-EDTA (1×), and fetal bovine serum (FBS), 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Sodium palmitate (PA), 3-(4,5-Dimethylthiazol-)2,5-diphenyl Tetrazolium Bromide (MTT), β-nicotinamide adenine dinucleotide (NADH), and tween-20 were purchased from Sigma Aldrich (Saint Louis, MO, USA). Sulfo-Nsuccinimidyl oleate (SSO) was purchased from Cayman Chemical (Ann Arbor, MI, USA). Dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), and nonfat dried milk were purchased from PanReac AppliChem (Glenview, IL, USA). Before each experiment, all solutions were freshly prepared. Absolute ethanol, hydrochloric acid, and methanol were purchased from Carlo Erba Reagents (Cornaredo, Milan, Italy).