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. 2016 Feb 22;14(2):e1002387. doi: 10.1371/journal.pbio.1002387

Fig 3. The rad50-V1269M allele impairs MRX association to a DSB in vivo.

Fig 3

(A, B) ChIP analysis. Exponentially growing YEPR cell cultures were transferred to YEPRG at time zero. Relative fold enrichment of the indicated fusion proteins at the indicated distances from the HO cleavage site was determined after ChIP with anti-HA or anti-Myc antibodies and subsequent qPCR analysis. Plotted values are the mean value with error bars denoting s.d. (n = 3). (C) Western blot with anti-HA and anti-Myc antibodies of extracts used for the ChIP analysis shown in (A) and (B), respectively. The same amount of extracts was separated on a SDS-PAGE and stained with Coomassie Blue as loading control. (D) Mre11-Myc was immunoprecipitated with anti-Myc antibodies and the immunoprecipitates were subjected to western blot analysis using anti-HA or anti-Myc antibodies. (E) As in (A), but showing Tel1-HA association to the DSB. (F) Western blot with anti-HA antibodies of extracts used for the ChIP analysis shown in (E). (G) Flag-tagged Tel1 (50 ng) was incubated with wild-type MRX or MRV1269MX (100 ng of each) and protein complexes were captured by anti-Flag beads. The supernatant (S) containing unbound proteins, the wash (W), and the eluate (E) fractions were analyzed by immunoblotting.