K666 100

Glucose consumption and lactate production rates were evaluated using commercial kits (BioVision, Mountain View, CA, USA; K666-100 and K627-100, respectively) according to the manufacturer’s instructions.

Glucose uptake was defined as the uptake of 2-deoxyglucose (2-DG) by HK-2 cells using a glucose uptake fluorometric assay kit (K666-100, BioVision, Milpitas, CA, USA) according to the manufacturer’s instructions. Generally, HK-2 cells were cultured in 96-well plates in keratinocyte SFM. When 50–60% confluent, the cells were treated with or without dapagliflozin (DAPA, 20 μM) for 48 h or cells were transfected with control siCON or siSGLT2 (200 nM) for 6 h. The cells were incubated in 0.5% bovine serum albumin (BSA)/phosphate-buffered saline (PBS) with or without 2-DG for 1 h. The relative fluorescence units (RFU) were measured at Ex/Em = 544/590 nm using a Fluoroskan Microplate Fluorometer (#5210470, Thermo Fisher Scientific, San Jose, CA, USA). The uptake of 2-DG in all samples was calculated by the 2-DG-6-phosphate (2-DG6P) standard curve and the RFU of samples.