Bolt 4 12 bis tris phosphate gels

WFA (>96% pure) was generously provided as a gift sample by 3P Biotechnologies Inc., (Louisville, KY, USA). A stock solution of WFA (5 mM) was prepared in dimethyl sulfoxide (DMSO) and stored in aliquots at −20 °C until use. MTT (3-[4,5-dimethylthiazol-2-yl]−2,5-diphenyltetrazolium bromide) was purchased from Alfa Aesar (Ward Hill, MA, USA). RIPA cell lysis buffer, BCA protein assay kit, PVDF membranes, phosphate–buffered- saline (PBS), ECL chemiluminescence reagent, and Bolt 4–12% Bis-Tris phosphate gels were purchased from ThermoFisher (Rockford, IL, USA). Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin and streptomycin were purchased from Life Technologies (Gibco, Grand Islands, NY, USA). Fetal bovine serum (FBS) was purchased from VWR, Seradigm Life Science (Radnor, PA, USA). The human recombinant TGFβ-1 (#8915) and TNFα (#8902) were purchased from Cell Signaling Technology (Danvers, MA, USA) and prepared as 50 µg/mL stock solutions in sterile 20 mM citrate, pH 3.0 and sterile PBS, respectively. The stock solution was stored in aliquots at −80 °C and diluted in serum-free DMEM before use.

A native mixture of Anthos (>95% pure) was isolated from a standardized highly enriched extract of bilberry using solvent extractions and C18 chromatography [24 ]. Bilberry (a European ‘cousin’ of blueberry) contains essentially the same anthocyanidin profile as blueberry. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) was purchased from Alfa Aesar (Ward Hill, MA, USA). RIPA cell lysis buffer, BCA protein assay kit, PVDF membranes, phosphate-buffered saline (PBS), ECL chemiluminescence reagent, and Bolt 4–12% Bis-Tris phosphate gels were purchased from ThermoFisher (Rockford, IL, USA). Dulbecco’s Modified Eagle’s Medium (DMEM), L-15 media, penicillin, and streptomycin were purchased from Life Technologies (Gibco, Grand Islands, NY, USA). Fetal bovine serum (FBS) was purchased from ThermoFisher (Rockford, IL, USA). The human recombinant TGFβ-1 (#8915) and TNFα (#8902) were purchased from Cell Signaling Technology (Danvers, MA, USA).