Anti glut3

Western blotting was performed as described previously [9] (link). Primary antibodies against CD9, CD63, CD81, HSP90, FGFBP1, SIPA1, THBS1, TGFBI, COL6A1, RPL10, SLC2A3 (GLUT3), MYO1D, RBP1, SMOC2, GLG1, and CEMIP (Protein-Tech Group, Rosemont, IL, USA) were used.
Western blotting was performed to verify the expression of exosome marker proteins and selected exoDEPs in SW620 and SW480 exosomes. Equivalent amounts of total protein (20 μg) were resolved by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the antibodies were diluted as follows: anti-CD9 (1:1000), anti-CD63 (1:1000), anti-CD81 (1:2000), anti-HSP90 (1:200), anti-FGFBP1 (1:500), anti-SIPA1 (1:2000), anti-THBS1 (1:500), anti-TGFBI (1:200), anti-COL6A1 (1:200), anti-RPL10 (1:1000), anti-GLUT3 (1:500), anti-MYO1D (1:200), anti-RBP1 (1:500), anti-SMOC2 (1:500), anti-GLG1 (1:1000), and anti-CEMIP (1:1000) (Protein-Tech Group, Rosemont, IL, USA). Proteins were detected using an enhanced chemiluminescence reagent (Santa Cruz Biotechnology, Dallas, TX, USA). Western blot signals were quantified using FluorChem E (Protein Simple, San Jose, CA, USA). All analyses were performed using western blots in at least two biological replicates.

The murine NIH-3T3 fibroblast cell line was obtained from the China Infrastructure of Cell Line Resources, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences. NIH-3T3 cells were grown in DMEM supplemented with 10% FBS (both Thermo Fisher Scientific, Inc.) at 37°C with 5% CO₂. The following antibodies were used in the present study: Anti-STYK1 (cat. no. 18028-1-AP), anti-GLUT1 (cat. no. 21829-1-AP), anti-GLUT2 (cat. no. 20436-1-AP), anti-GLUT4 (cat. no. 21048-1-AP), anti-hexokinase (HK)1 (cat. no. 19662-1-AP), anti-platelet phosphofructokinase (PFKP) (cat. no. 13389-1-AP) and anti-pyruvate kinase (PKM)1 (cat. no. 15821-1-AP), all from ProteinTech Group, Inc.; anti-GLUT3 (cat. no. ab191071) and anti-pyruvate dehydrogenase α1 (PDHA1) (cat. no. ab168379), all from Abcam, Inc.; anti-β-actin (cat. no. 4970) and anti-β-tubulin (cat. no. 2146), both from Cell Signaling Technology, Inc.; and HRP-conjugated secondary antibody (cat. no. TA130003) from OriGene Technologies, Inc. MTT, DMSO and G418 were purchased from Sigma-Aldrich; Merck KGaA.