biostudies-otherUnknownJeroen PenningsHorizon Europe Research and Innovation ProgrammeEuropean Union’s Horizon 2020 Research and Innovation ProgrammeHomo sapiens (human)https://www.ebi.ac.uk/biostudies/studies/S-BSST2297biostudies-other101057014964537Jeroen PenningsfalseComet assay in HepG2 cells after compound exposureTo 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. The 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. This study was performed as part of the RISK-HUNT3R project and the PARC project.2026-03-31T00:00:00Z2026-06-29T04:01:37.767Z2025-11-20T14:35:05.631ZS-BSST2297