Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells - Université de Montpellier
Article Dans Une Revue STAR Protocols Année : 2023

Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells

Résumé

Translesion DNA synthesis (TLS) is an evolutionarily conserved branch of the cellular DNA damage tolerance pathway that is often exploited by cancer cells to overcome therapy resistance. Here, we present a protocol to analyze endogenous TLS in single mammalian cells in the absence or presence of DNA damage. We describe steps for detecting chromatin-bound TLS factors, such as monoubiquitinated PCNA(mUb) and TLS DNA polymerases (pols) by flow cytometry. We then detail a procedure to detect their nuclear localization using immunofluorescence.
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Dates et versions

hal-04289638 , version 1 (16-11-2023)

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Tom Egger, Antoine Aze, Domenico Maiorano. Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells. STAR Protocols, 2023, 4, ⟨10.1016/j.xpro.2023.102361⟩. ⟨hal-04289638⟩
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