Anti glut2

After treatment with glucose-free DMEM for 3 h, STC-1 cells were incubated with different concentrations of glucose (0, 5.6, 25, 100, and 200 mM) for 1 h. The cells were harvested, and cell lysates were prepared. BCA Protein Assay Kit (Beyotime, Jiangsu, China) was used to determine the protein content in the lysates. For Western blot analysis, 50-100 μg of protein from each sample was subjected to separate on a SDS-PAGE gel. The blots of proteins in polyvinylidene difluoride membranes (Merck Millipore, Darmstadt, Germany) were incubated with the appropriate primary antibody and incubated with horseradish peroxidase (HRP)-conjugated secondary antibody. The blots were detected with chemiluminescence (ECL-kit, Beyotime, Jiangsu, China) followed by autoradiography. The antibodies used for Western blot were as follows: anti-SGLT1 (Abcam, Cambridge, UK), anti-GLUT2 (Abcam, Cambridge, UK), anti-STR subunit TAS1R2 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-STR subunit TAS1R3 (Santa Cruz Biotechnology, Santa Cruz, CA), and anti-β-actin antibody (Santa Cruz Biotechnology, Santa Cruz, CA). Quantification of protein bands was performed using ImageJ software version 1.42 (National Institutes of Health, Bethesda, MD, USA).