Western Blot and IHC Analysis of Key Proteins

GLUT1 (12939S, used for Western blotting), HSP90 (4877S), p53 (2524S), Cleaved Caspase-3 (9661S) and GAPDH (2118S) antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA), and p21 (ab7960) antibody was purchased from Abcam (Cambridge, MA, USA). GLUT1 (ab652) was used for IHC at a concentration of 1:500 and was purchased from Abcam (Cambridge, MA, USA). Normal rabbit IgG was used as a negative control. For Western blotting, 50–100 µg of whole cell lysate was resolved over SDS PAGE gels using pre-cast 10% NuPAGE Bis-Tris gels (Thermo Fisher Scientific) and transferred onto PVDF membranes (IPVH00010, pore size: 0.45 µm) (Millipore Sigma) prior to analysis. Horseradish peroxidase-conjugated secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). IHC analysis was performed as described [7 (link)].