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. 2013 Sep 17;41(22):10371–10378. doi: 10.1093/nar/gkt820

Figure 2.

Figure 2.

The genome integrity checkpoint is pre-activated in ixr1 mutants and is activated further in response to DNA-damaging drugs and replication block. (A) Western blot analysis of Rnr3 and Rnr4 levels in wild-type (wt) (TOY632) and ixr1Δ (TOY732) strains before and after 2-h treatments with 0.2 mg/l 4-NQO, 0.02% MMS, 200 mM HU, 2.5 mMcisplatin or 325 μM t-BHP. Protein levels were quantified in relative units (RU, levels of Rnr3 or Rnr4 divided by the level of tubulin in the same sample lane as described in ‘Materials and Methods’ section). ND: not detected. (B) Western blot analysis of Rnr3, Rnr4 and Sml1 levels in wild-type (wt) (TOY632) and ixr1Δ (TOY732) strains. Cells were synchronized with α-factor, released into fresh YPD media and collected at the indicated time points. Upper panel: corresponding flow cytometer charts. as: asynchronous culture. RU, relative units; ND, not detected.