Figure 1: HK1 interacts with c-Src.
From: c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis
(a) HEK 293T cells were co-transfected with 2 μg of HA-c-Src and equal amount of each of plasmids expressing Flag-tagged enzymes involved in glycolysis (hexokinase 1, HK1; phosphoglucose isomerase, PGI; phosphofructokinase-1, PFK-1; aldolase; triose phosphate isomerase, TPI; glyceraldehydes-3-phosphate dehydrogenase, GAPDH; phosphoglycerate kinase 1, PGK1; phosphoglycerate mutase 1, PGM1; enolase; pyruvate kinase M2, PKM2). Immunoprecipitation (IP) were performed with HA antibody after 24 h of transfection. WB, western blot, TCL, total cell lysate. (b,c) HEK 293T cells were transfected with HA-c-Src and Flag-HK1 in combinations as indicated. Reciprocal IPs were carried out to precipitate Flag-HK1 (b) and HA-c-Src (c). (d) Endogenous c-Src in lysate of HCT116 cells was precipitated with anti-c-Src, followed by WB to detect c-Src and HK1. (e) GST pull down was performed with His-HK1 and GST-c-Src, followed by coomassie brilliant blue staining (left panel) and WB with HK1 antibody for His-HK1 and GST antibody for GST-c-Src. (f) HeLa cells were co-transfected with Flag-c-Src and HA-HK1. After 24 h of transfection, immunofluorescence staining was performed to observe the co-localization of c-Src and HK1. Scale bars, 30 μm.
