L6 cells were seeded in a 96-well transparent plate with low density in 100 µL DMEM. After 6 days of differentiation, as described above. L6 myotubes cells were treated with B4 (64, 16, 4 μg/mL), and PBS buffer dissolved in 100 μL DMEM medium, six wells were repeated in each group. After 24 h of treatment, the cell supernatant was taken, and the glucose concentration was detected by the glucose oxidase method using an automatic biochemical analyzer (HITACHI, Japan). Glucose consumption in the culture medium reflects intracellular glucose uptake.
L6 cells were cultured in a 6-well transparent plate. There were four wells in the normal group and the B4-treated groups. After B4 (64, 16, 4 μg/mL) treatment for 24 h, cells were collected and the intracellular glucose was lysed according to the instructions of the glucose content assay kit (APPLYGEN, Beijing, China), the supernatant of lysate was used to measure the glucose content using the oxidase method, and the protein in the precipitate was detected by a BCA kit (Beyotime, Beijing, China). The amount of glucose per gram of protein was calculated and deemed as glucose uptake.
L6 cells were cultured in a 6-well transparent plate. There were four wells in the normal group and the B4-treated groups. After B4 (64, 16, 4 μg/mL) treatment for 24 h, cells were collected and the intracellular glucose was lysed according to the instructions of the glucose content assay kit (APPLYGEN, Beijing, China), the supernatant of lysate was used to measure the glucose content using the oxidase method, and the protein in the precipitate was detected by a BCA kit (Beyotime, Beijing, China). The amount of glucose per gram of protein was calculated and deemed as glucose uptake.