Hamamelitannin

Full extracts (60% ethanol) of Hamamelis virginiana L. leaf or bark, as well as ultrafiltration (UF) concentrates and filtrates were prepared as previously described [47] (link). For bark extract fractionation into UF-concentrate and filtrate, a Pro Flux M12 Tangential Flow Filtration System (Millipore) with Pellicon 2 Ultrafiltration Cassettes (C Screen, Millipore) was used. All extracts and UF-fractions were prepared at Dr. Willmar Schwabe GmbH & Co. KG, Karlsruhe and provided as dry powders. Concentrations indicated designate dry extract weight per volume of solvent. Gallic acid monohydrate (“gallic acid”), penta-O-galloyl-β-D-glucose (“pentagalloylglucose”), hamamelitannin, tannic acid and (−)-epigallocatechin gallate (EGCG) were purchased from Sigma-Aldrich. The buffer capacity of the cell culture medium fully neutralized the acidity of compounds added to in vitro experiments.

Antibodies against AMPKα and phospho-AMPK α (p-AMPKα) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against proliferator-activated receptor gamma coactivator 1-α (PGC-1α), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), heat shock protein 90 (HSP90), and translocase of outer membrane 20 (TOM20) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Furthermore, antibodies against CV-ATP5A, CIII-UQCRC2, CII-SDHB, and C1-NDFUB8 were purchased from Invitrogen (Carlsbad, CA, USA). LDHA antibody was supplied by Abcam (Cambridge, MA, USA). The 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) were purchased from Merck (Darmstadt, Germany). The gallic acid, catechin, hamamelitannin, ellagic acid, protocatechuic acid, and ziuyuglycoside I was supplied by Sigma-Aldrich (St. Louis, MO, USA).

Hamamelitannin (HAM), vancomycin (VAN), cefoxitin (CFX), cefazolin (CZ), cefalonium (CL), cephalexin (CFL), linezolid (LNZ) and fusidic acid (FA) were purchased from Sigma Aldrich (Bornem, Belgium). Tobramycin (TOB) and daptomycin (DAP) were purchased from TCI (Tokyo, Japan). HAM was stored in DMSO at −20 °C. All antibiotics were dissolved in ultrapure water (with the exception of CZ, CL and CFL, which were dissolved in phosphate buffer). HAM was used at 250 μM concentrations unless otherwise mentioned.