Differentiated C2C12 cells were incubated with conditioned medium obtained from the RINm5f cells seeded onto the PDA-PLGA scaffolds for 24 h. Cells were then cultured in FBS-free DMEM (15 mM D-glucose) supplemented with 0.2% BSA for 48 h. The media glucose concentration was determined as described above. Glucose consumption was calculated based on the difference between the initial glucose concentration and the residual glucose concentration of the culture medium. For glucose uptake determination, cells were washed three times with KRB and incubated with conditioned medium or normal medium for 10 min, and 100 μM 2-NBDG (Invitrogen, United States) was added to the medium for 30 min. The medium was removed, and the cells were then washed twice with ice-cold PBS. The fluorescence intensity in each well was then measured at 485 nm/535 nm (excitation wavelength/emission wavelength) using an Epoch fluorescence microplate reader (Biotek, United States). Cells cultured without conditioned medium were defined as the control group (Con), cells cocultured with conditioned medium were defined as the RINm5f group (RINm5f).
Epoch fluorescence microplate reader
Manufactured by Agilent Technologies
Sourced in United States
The Epoch fluorescence microplate reader is a laboratory instrument designed for quantitative fluorescence measurements. It can detect and measure fluorescent signals from microplate samples. The core function of the Epoch is to provide accurate and reliable fluorescence detection capabilities for various applications.
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2 protocols using epoch fluorescence microplate reader
1
Glucose Uptake in Differentiated C2C12 Cells
2
Glucose Uptake in Differentiated C2C12 Cells
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Differentiated C2C12 cells were incubated with miR-27a plasmid in the absence or presence of 20 μM rosiglitazone for 48 h. Cells were then cultured in FBS-free DMEM (15 mM D-glucose) supplemented with 0.2% BSA for 48 h. The Glucose concentration in the medium was determined as above. Glucose consumption was calculated from the difference between the initial glucose concentration and the residual glucose concentration of the culture medium. For glucose uptake determination, cells were washed three times with KRB, and incubated with or without 100 nM insulin in glucose-free DMEM for 10 min, and 100 μM 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG; Invitrogen, USA) was added to the medium for 30 min. The medium was removed, and the cells then were washed twice with ice-cold PBS. The fluorescence intensity in each well was then measured at 485 nm/535 nm (excitation wavelength/emission wavelength) using an Epoch fluorescence microplate reader (Biotek, USA).
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