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Journal: bioRxiv
Article Title: Resolution of collapsed forks is separate from completion of DNA synthesis
doi: 10.1101/2025.03.03.641087
Figure Lengend Snippet: (A) Cartoon depicting formation of full length molecules by synthesis through the LacR array or independent of LacR array displacement. (B) Leading collapse was induced as depicted in . Samples from T=120 min were then purified and analyzed by restriction digest using either AlwNI alone or SacI and KpnI combined. Digested samples were separated on an agarose gel and visualized by autoradiography. (C) Quantification of full length linear products that arose from AlwNI digestion in (b). Mean ± S.D., n = 3 independent experiments. Data were analyzed by unpaired t-test. (D) Quantification of linearized lacO array fragments that arose from SacI-KpnI digestion in (b). Mean ± S.D., n = 3 independent experiments. Data were analyzed by unpaired t-test. (E) Cartoon depicts generation of circular monomers (CMs) by extensive degradation of seDSBs. (F) Leading collapse was induced as depicted in in mock- or MRE11-immunodepleted extracts. Samples from were separated on an agarose gel and visualized by autoradiography. (G) Quantification of σ structures (σs) from (f). Mean ± S.D., n = 3 independent experiments. (H) Quantification of nicked plus supercoiled circular monomers (CMs) from (f). Mean ± S.D., n = 3 independent experiments. (I) Cartoon depicts the retention of σ structures (σs) in response to extensive degradation of a lagging seDSB due to resynthesis of lagging strands. (J) Lagging collapse was induced as depicted in . in the presence of either vehicle or BRC peptide. Samples from were separated on an agarose gel and visualized by autoradiography. (K) Quantification of high molecular weight products (HMw) from (i). Mean ± S.D., n = 3 independent experiments.
Article Snippet:
Techniques: Purification, Agarose Gel Electrophoresis, Autoradiography, High Molecular Weight