C2C12 cells treated with the CMs were washed with PBS and starved in glucose-free DMEM containing 2% FA-free FBS for 3 h. They were subsequently incubated in the presence of insulin (100 nM) for 10 min, then washed with ice-cold Krebs buffer and 2-NBDG (Cayman, USA) (150 μg/mL) was added into the culture medium for 30 min at 37 °C. The 2-NBDG fluorescence intensity was measured using a microplate reader at an excitation wavelength of 480 nm and an emission wavelength of 540 nm.
In addition, 10 ng anti-resistin antibody (ab136877, Abcam, USA) was added into the obtained 20 mL CM5 (CM5-AntiR). Then, the C2C12 cells were cultured with the mixed medium. Glucose uptake was assessed using the 2-NBDG fluorescence intensity. The CMM value for the glucose uptake was set as 100%.
In addition, 10 ng anti-resistin antibody (ab136877, Abcam, USA) was added into the obtained 20 mL CM5 (CM5-AntiR). Then, the C2C12 cells were cultured with the mixed medium. Glucose uptake was assessed using the 2-NBDG fluorescence intensity. The CMM value for the glucose uptake was set as 100%.