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. 2020 Mar 31;21(7):2399. doi: 10.3390/ijms21072399

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Establishment and validation of RUES2 and differentiation of cardiomyocytes in vitro. (A) Schematic of the RUES2-CMs differentiation protocol. (B) Flow cytometric analysis of pluripotent stem cell markers expression: Oct-4, SSEA-4, Nanog, and a positively selected cardiac marker (cTnT). (C) Representative fluorescent images of pluripotent stem cell markers Oct-4 and cardiac markers: α-actinin and cTnT. Scale bar: 50 μm (upper panel); 20 μm (lower panel). (D) PD-L1 expression on RUES2-CMs by flow cytometry and (E) immunofluorescence staining, scale bar: 25 μm. The quantitative result as the right panel. *** p < 0.001 versus control (n = 3), one-way ANOVA, posthoc Bonferroni test. RUES2, Rockefeller University embryonic stem cell line 2; cTnT, cardiac troponin T; Oct4, octamer-binding transcription factor 4; PD-L1, programmed death-ligand 1; RUES2-CMs, Rockefeller University embryonic stem cell line 2-cardiomyocytes.