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After centrifuging the plates for 5 minutes at 1,500 rpm, duplicate wells were harvested and lysed with 350 l of RLT buffer

To confirm that the overexpression of HRASV12 and cyclin E1 via retroviral transduction induced R-loop formation, we isolated genomic DNA from U2OS cells transduced with the respective retroviral vector and performed slot blot analysis using the S9.6 antibody, which recognizes RNA:DNA hybrids