2 7 dichlorodihydrofluorescein diacetate dcfda

Primary glomerular microvascular endothelial cells (GMVEC), purchased from Cell Systems (Kirkland, WA, USA), were cultured in complete classic medium (#4Z0-500) supplemented with culture boost. The passage number was used between 3 to 7. This primary cell line was utilised as any other relevant cell lines are immortalised rather than human primary cells and were therefore not appropriate. Endothelial growth factor (VEGF-A₁₆₅) was purchased from Thermo-Fisher (Paisley, UK). Pure and analytical grade artificial sweeteners, aspartame, saccharin, and sucralose, the CCK-8 cell viability kit, FITC-dextran, N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), methanol, and ethyl acetate were purchased from Sigma-Aldrich (Dorset, UK). Lactisole, a sweet taste inhibitor, was purchased from Cayman Chemical (Ann Arbor, MI, USA). The cAMP-Screen Direct System kit and GSH Bioxytech activity kit were purchased from Applied Biosystems and Merck Millipore respectively. DharmaFECT™ reagent and siRNA (T1R3 and non-specific, scrambled) were purchased from Dharmacon (Cambridge, UK). Anti VE-cadherin and fluorescent secondary antibodies, deuterated sucralose, and sucralose-D6 (SC-220145) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 2,7-dichlorodihydrofluorescein diacetate (DCFDA) was purchased from Abcam (Cambridge, UK).

Ferulic acid was purchased from Sigma-Aldrich Co. LLC (CAS:537-98-4; St. Louis, MI, USA), and 2’,7’-dichlorodihydrofluorescein-diacetate (DCF-DA) was from Abcam (Cambridge, UK). Fetal bovine serum (FBS) was purchased from HyClone Laboratories (Logan, UT, USA). Unless specified otherwise, other chemicals and laboratory consumables were purchased from Sigma-Aldrich Co. LLC and Falcon Labware (BD Biosciences, Franklin Lakes, NJ, USA), respectively. C57BL/6 mice (6 weeks old) were purchased from Damul Science (Daejeon, Republic of Korea) and equilibrated for 7 days before use. During the experimental period, all mice were housed at 22 ± 1 °C and 55 ± 5% humidity, along with 12 h light/dark autocycle, allowing ad libitum feeding in the Animal Center of School of Dentistry, Jeonbuk National University.