Synthesis and Characterization of DPP Probes

DMEM GlutaMAX (Gibco), Fetal Bovine Serum (FBS; Gibco/Life Technologies, Qualified US origin or Gemini Benchmark^™ 100-106), Live Cell Imaging Solution (Molecular Probes), Opti-MEM (Gibco), Lipofectamine 3000 reagent (Invitrogen), Lipofectamine RNAiMAX transfection reagent (Invitrogen), polyethylenimine (PEI) (Sigma; average Mw ~25,000 by LS, average Mn ~10,000 by GPC, branched), Dynabead Protein G magnetic beads (Invitrogen) MitoTracker Deep Red FM (Invitrogen), Hoechst 33342 (Fisher), 2-BP (Sigma), MitoPY1 (Sigma), PY1 (Sigma), Palmostatin B (PalmB) (EMD Millipore), Charcoal-filter FBS (A3382101, Thermo Fischer), MTS (BioVision), Phenazine methosulfate (Sigma) were purchased as mentioned in parenthesis. siRNAs targeting human ABHD10 (SI04229519), human APT1/LYPLA1 (SI03246586) and human PRDX5 (SI00096971, SI02638888, SI02638902), as well as non-targeting (NT) control siRNA (SI03650325), were purchased from Qiagen. Silica gel P60 (40-63 μm, 230-400 mesh; SiliCycle) was used for column chromatography. Analytical thin layer chromatography was performed using precoated 60 F254 silica gel sheets (0.25 mm thick; SiliCycle). DPP-2^{30 (link)}, DPP-5^{32 (link)} and mitoDPP-2^{20 (link)} were synthesized as previously reported. All chemicals for synthesis were purchased from Sigma-Aldrich or Fisher Scientific and used as received. ML348 was purchased from Tocris (Bristol, UK).