Anti glut1 sc 377228

Both HepG2 and Huh7 cells were transfected with control miRNA or miR-505 mimics. After transfection for 48 h, cells were harvested and lyzed with the RIPA buffer (Beyotime, Shanghai, China) on ice for 10 min. The protein concentration was determined via the BCA protein assay (Beyotime, Shanghai, China). Equal amount of protein was separated by SDS/PAGE and transferred on to the nitrocellulose membranes (Millipore, Bedford, MA, U.S.A.). After blocking with 5% non-fat milk for 1 h at RT, membrane was incubated with the primary antibody (anti-IGF-1R (9750S), anti-pAKT (Ser⁴⁷³, 9271S), anti-AKT (9272S), Cell Signaling Technology, Danvers, MA, U.S.A.; anti-GLUT1 (sc-377228), Santa Cruz Biotechnology (Dallas, TX, U.S.A.) for 2 h at RT. Afterward, membrane was incubated with horseradish peroxidase (HRP)–conjugated secondary antibody (1:3000, Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) for 1 h at RT. Protein bands were visualized with Enhanced ECL detection kit (Millipore, Bedford, MA, U.S.A.).