Fig. 2. Glycolytic inhibiting activities of Compd 27.

(A and B) Antiproliferation activity of Compd 27 or 3-BP (the known standard control as the HK2 inhibitor) against glioma U87 cells (n = 6). (C) U87 cell apoptosis triggered by Compd 27 with flow cytometry analysis. Cell staining by annexin V–FITC/PI. Statistical significance was calculated using the unpaired two-tailed Student’s t test. ***P < 0.001 (n = 3). (D) Fluorescent images of U87 cells treated by Compd 27 and 3-BP. Cell nuclei–stained by 4′,6-diamidino-2-phenylindole (DAPI) (blue); cytoskeleton stained by phalloidine-FITC (green). Scale bar, 100 μm. (E) Schematic representation of the binding structures of Compd 27-HK2 complex predicted by Glide XP docking simulations. (F) Two-dimensional schematic diagram of the binding patterns of Compd 27-HK2 complex, highlighting hydrogen bonds and dominant hydrophobic interactions. Color codes: pink (polar amino acids); green (greasy residues). (G) Differences of the interaction scores (Eint_{Compd 27} − Eint_3-BP) between Compd 27 and 3-BP on a per-residue basis with the important residues highlighted. Calculations of total interaction energy, van der Waals interaction, and hydrogen bonding scores between the ligands and the protein residues by Glide module in Schrödinger.