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. 2021 Aug 20;9(8):1057. doi: 10.3390/biomedicines9081057

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© 2021 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 (https://creativecommons.org/licenses/by/4.0/).

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Flow cytometry detection of DNA damage. To evaluate DNA damage, we used the H2A.X phosphorylation assay, a cell-based assay using flow cytometry detection of levels of phosphorylated histone H2A.X on serine 139 (γ-H2AX), formed in response to DNA double-strand breaks (DSBs) generated by exogenous genotoxic agents. Cells for each condition were treated in vitro with 0, 0.5, and 1 μmol/L cisplatin for 72 h. See the Materials and Methods section for more details. RDM siRNA NEG = siRNA scramble-treated HepG2 cells; RDM siRNA PER1 = siRNA PER1-treated HepG2 cells; M1 siRNAneg = siRNA scramble-treated macroH2A1 knocked down HepG2 cells; M1 siRNA PER1 = siRNA PER1-treated macroH2A1 knocked down HepG2 cells; three biological replicates were each assayed in triplicate.