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. 2014 Dec 23;34(3):326–343. doi: 10.15252/embj.201489184

Figure 5.

Figure 5

USP45 knockout cells are hypersensitive to DNA damage
  1. Clonogenic survival assays were carried out in wild-type (WT), USP45 knockout (KO) or USP45 KO cells re-expressing wild-type USP45 (RESWT) or catalytically inactive USP45 [Cys199Ala] (RESCA). The cells were treated with the indicated doses of mitomycin C (MMC) 16 h. Medium was changed and number of colonies quantified after 10 days. Each data point is the average of 3 experiments undertaken with 6 replicates ± SD.
  2. Control immunoblot analysis demonstrating that in the USP45 KO U2OS rescue cell lines, similar levels of wild-type and catalytically inactive USP45 [Cys199Ala] were expressed.
  3. As above, except rescue experiments are undertaken using the non-ERCC1-binding USP45 [Asp25Ala, Glu26Ala] mutant.
  4. As in (A), except that DNA damage is induced using UV-C irradiation (E) or 16 h hydroxyurea treatment (F). Similar results were obtained in at two separate experiments.