{"database":"biostudies-other","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Jeroen Pennings"],"funding":["Horizon Europe Research and Innovation Programme","European Union’s Horizon 2020 Research and Innovation Programme"],"species":["Homo sapiens (human)"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-BSST2297"],"repository":["biostudies-other"],"funding_grant_id":["101057014","964537"],"pubmed_authors":["Jeroen Pennings"],"additional_accession":[]},"is_claimable":false,"name":"Comet assay in HepG2 cells after compound exposure","description":"To quantify oxidative DNA damage, we performed the comet assay on maturated HepG2 cells after 24 h of compound exposure. This was done using cells receiving repeated exposure, as well as cells receiving repeated exposure and recovery. \nThe enzyme-modified alkaline comet assay was performed as described in [Collins et al. Nat Protoc. 2023;18(3):929–989]. This was done in the presence or absence of the enzyme DNA-formamidopyrimidine glycosylase (Fpg). At least 50 comets per gel were scored to obtain the median %DNA tail intensity (TI) and the mean of two replicate gels was calculated per sample. The results obtained after incubation without Fpg-enzyme only represent the DNA strand breaks and alkali-labile sites. The difference between the samples treated with Fpg-enzyme and those incubated without was calculated for each sample to derive the net Fpg-sensitive sites (%TI), which represent DNA oxidation. A positive assay control was included by using aliquots of frozen Hela cells that were exposed to 2mM KBrO3 for 1 hour (generating an oxidized DNA substrate) in the same batch analysis. The net Fpg results were normalized using this assay control to correct for inter-assay variation.\nThis study was performed as part of the RISK-HUNT3R project and the PARC project.","dates":{"release":"2026-03-31T00:00:00Z","modification":"2026-06-29T04:01:37.767Z","creation":"2025-11-20T14:35:05.631Z"},"accession":"S-BSST2297","cross_references":{}}