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. 2017 Aug 16;36(19):2829–2843. doi: 10.15252/embj.201796717

Figure 1. Whole‐genome mapping of UV‐induced DNA lesions.

Figure 1

  1. Average pixel volumes of slot blot analysis using purified DNA from IMR90 cells treated with indicated UVC doses. Standard deviation of three technical replicates is shown.
  2. Illustration of experimental method. Briefly, IMR90 primary cells were irradiated with 100 J/m2 UVC for <10 s, cells immediately lysed and DNA purified. Cyclobutane pyrimidine dimer (CPD)‐containing fragments were immunoprecipitated (IP'ed) followed by repair of CPDs and 6–4 photoproducts (6–4 PPs) with photolyases. DNA photoproducts were then sequenced via Illumina sequencing.
  3. Slot blot of eluate from mock IP (−Ab) and CPD‐IP (+Ab) using CPD antibody.
  4. Slot blot using CPD antibody of mock (−photolyase, PL) and repaired (+PL) IP'ed DNA irradiated with either 100 or 500 J/m2 UVC. Repair efficiency was determined to be greater than 95%.
  5. Pearson's correlation between sequencing replicates with indicated bin sizes.
  6. Boxplots indicating frequency of indicated pyrimidine dimers within sequenced reads. Middle quartiles are represented by boxes, top and bottom quartiles are represented by whiskers. IP replicates are merged. Wilcox test indicates that TpT and TpC are statistically enriched (P‐value < 0.0001) in IP versus input.