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. 2011 Dec 15;7(12):e1002417. doi: 10.1371/journal.pgen.1002417

Figure 6. Rif1 does not associate with an HO-induced DSB. Strains were grown overnight on raffinose; galactose was added to the medium at time 0 and samples collected every 2 hours.

Figure 6

Galactose induces GAL-HO-nuclease to cut an unique DSB at the MATa locus [7]. Although a DSB is usually repaired by recombination, recombination was prevented in JKM139-derived strains by deletion of the donor locus [7]. (a–b) Chromatin immuno-precipitation experiments indicating the association of RPA and Ddc2 with chromatin at 0.2 kb right from the DSB, in RIF1+ (white columns) and rif1Δ (grey columns) JKM139-derived strains. (c–d) As in a–b, except that the association of RPA and Ddc2 with chromatin at 2 kb right from the DSB was analyzed. e) Chromatin immuno-precipitation with Myc antibody in the Rif1-Myc JKM139-derived strain. Legend indicates the time in galactose. The numbers on the X-axis indicate the location of the DSB (marked as 0) and the distance from the DSB (in kilobases). (f) As in (e), except that immuno-precipitation with Myc antibody was performed in the untagged JKM139 strain.