Glucose uptake assay
MTT assay was first performed using Vybrant MTT Cell Proliferation Assay
Kit (Invitrogen, Carlsbad, CA, USA) and manufacturer’s instructions to
find HSG4112 concentration range for cell viability. Glucose uptake
assay on differentiated 3T3-L1 adipocytes and C2C12 myotubes were
performed in accordance with the manufacturer’ instructions in Glucose
Uptake Assay Kit (ab136955, Abcam Inc., Cambridge, UK). DMEM/F-12 media
(Gibco BRL, Grand Island, NY, USA) and KRPH buffer (BSA, Sigma-Aldrich
Co., St. Lousi, MO, USA) were purchased separately.
2-Deoxy-D-glucose (2DG) – 2-hydroxyl group replaced by
hydrogen so that it cannot undergo further glycolysis – was added and
2-deoxyglucose-6-phosphate (2DG6P) was measured as glucose output.