HeLa-S3 (ATCC, purchased and authenticated by STR analysis in 2016) and HeLa GFP-RAD52 (this study) were cultured in DMEM (Sigma-Aldrich) supplemented with 10% FCS (Sigma-Aldrich or BioSell) and 1% NEAA (Merck Millipore). Immortalised human primary fibroblast 82–6 hTERT (WT)36 (link), HSC-62 hTERT (BRCA2*)21 (link) and 2BN hTERT (XLF*)37 (link) cells were cultured in MEM (Sigma-Aldrich) with 20% FCS and 1% NEAA. U2OS EJ2-GFP reporter cells24 (link) were cultured in 2 μg/ml puromycin DMEM, supplemented with 10% FCS and 1% NEAA. WT and KO U2OS cell lines (RAD52, POLθ, RAD52/POLθ)26 (link) were cultured in DMEM (Sigma-Aldrich) supplemented with 10% FCS (Sigma-Aldrich or BioSell), 1% Glutamine (Gibco). Cells were routinely tested for mycoplasma contamination by PCR (Minerva Biolabs). All cell lines were maintained at 37 °C in a 5% CO2 incubator.
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Cell Culture Protocols for DNA Repair Studies
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Corresponding Organization :
Other organizations : Technical University of Darmstadt, University of California, Davis, The University of Texas MD Anderson Cancer Center
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Variable analysis
independent variables
- Cell lines used: HeLa-S3, HeLa GFP-RAD52, immortalised human primary fibroblast 82–6 hTERT (WT), HSC-62 hTERT (BRCA2*), 2BN hTERT (XLF*), U2OS EJ2-GFP reporter cells, WT and KO U2OS cell lines (RAD52, POLθ, RAD52/POLθ)
- Not explicitly mentioned
- Culture conditions: DMEM (Sigma-Aldrich) supplemented with 10% FCS (Sigma-Aldrich or BioSell) and 1% NEAA (Merck Millipore) for HeLa-S3, HeLa GFP-RAD52, WT and KO U2OS cell lines
- Culture conditions: MEM (Sigma-Aldrich) with 20% FCS and 1% NEAA for immortalised human primary fibroblast 82–6 hTERT (WT), HSC-62 hTERT (BRCA2*), and 2BN hTERT (XLF*) cells
- Culture conditions: 2 μg/ml puromycin DMEM, supplemented with 10% FCS and 1% NEAA for U2OS EJ2-GFP reporter cells
- Mycoplasma contamination testing by PCR (Minerva Biolabs)
- Incubator conditions: 37 °C in a 5% CO2 incubator
- No positive or negative controls explicitly mentioned
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