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. 2020 Jan 8;19(3):1701–1710. doi: 10.3892/etm.2020.8436

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Copyright: © Ke et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 4. — ROCK inhibitors prevent the apoptosis of dissociated hiPSC-CMs in serum-free medium. (A) Following treatment with or without fasudil for 72 h, cells were collected and analyzed by flow cytometry using Annexin V-FITC/PI staining. ****P<0.0001. (B-E) hiPSC-CMs were cultured for 3 days in the presence or absence of Y27632. (B) Flow cytometric analysis were used to detect apoptosis. **P<0.01. (C) The expression of Bcl-2, Bax and caspases 3 and 8 were detected by reverse transcription-PCR and (D) quantitative PCR. ***P<0.001. (E) The expression of cleaved caspase-3 and caspase-8 were analyzed by western blotting, with GAPDH as the loading control. ****P<0.0001. (F) Y27632 treatment reduced the activity of caspase-3 in detached hiPSC-CMs in suspension culture. **P<0.01. ROCK, Rho-associated kinase; hiPSC-CMs, human induced pluripotent stem cells-derived cardiomycytes; PI, propidium iodide; PNA, p-nitroaniline; OD, optical density.