Hironori Hara et al. BTS 2018;3:

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Hironori Hara et al. BTS 2018;3:639-653 Generation of the Novel Fluorinated Compound TT-10 (A) Chemical structures of the 26 analogues of TAZ-12. The chemical structure modified is highlighted with a colored circle or square. (B) Chemical structures of TAZ-12 and TT-10. (C) Ratios of 5-ethynyl-2ʹ-deoxyuridine (EdU)–positive cardiomyocytes (CMs) and mean CM size 40 h after treatment with the indicated concentrations of TT-10. n = 7 per group. (D) Relative cell number of CMs 40 h after treatment with TT-10 (10 μmol/l), relative to untreated CMs. n = 12 per group. (E) Ratios of EdU-positive CMs after the treatment with indicated reagents at the following concentrations: TAZ-12, 10 μmol/l; TT-10, 10 μmol/l; NRG1, 100 ng/ml; and acidic fibroblast growth factor (FGF1), 100 ng/ml. Representative images of TAZ-12– and TT-10–treated CMs are shown. The relative values are normalized to the untreated control. Arrowheads indicate positive CMs. n = 4 per group. Scale bar: 100 μm. (F and G) Percentages of pH3-positive (F) and Aurora B-positive (G) CMs after treatment with TAZ-12 and TT-10. Representative images are shown. Arrowheads indicate positive CMs. n = 8 (F) and n = 4 (G) per group. Scale bar: 100 μm in A; 50 μm in B. (H) MTS cell viability assay. The positive control was 100 μmol/lH2O2. n = 3 per group. (I) Effects of TAZ-12 and TT-10 on proliferative activities of non-CMs. Rat cardiac fibroblasts, NIH3T3 fibroblasts, and human umbilical vein endothelial cells (HUVECs) were cultured for 24, 18 and 18 h, respectively. n = 5 (rat cardiac fibroblasts), n = 7 (NIH3T3 fibroblasts), and n = 5 (HUVECs) per group. (J) Percentages of mono- or binucleated CMs. Representative images are shown. CMs infected with Fucci-expressing adenoviruses under the Tnnt2 promoter were treated with TAZ-12 (10 μmol/l) and TT-10 (10 μmol/l) for 40 h. n = 4 per group. Scale bar: 100 μm. *p < 0.05, **p < 0.01, and ***p < 0.001 versus the untreated control. Hironori Hara et al. BTS 2018;3:639-653 2018 The Authors