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. 2005 Jan 15;19(2):255–269. doi: 10.1101/gad.321105

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Figure 4. — Formaldehyde-independent hotspot association of Spo11 precedes DSB formation in rad50S. (A) The left upper panel (IP after FA cross-linking) shows specific enrichment of the PP1 product. The panel below (IP w/o FA) shows a very similar enrichment, although the signal appears slightly later. (WCE) Whole-cell extract. The Western blot shows significant expression of Spo11-Myc from the 2-h time point onward. The Southern blot shows signal corresponding to DNA fragments generated by meiotic DSBs at the his4XLEU2 hotspot from the same time course. (B) qPCR of rad50S ChIP-DNA (IP–FA and +FA cross-linking) at the YCR047c hotspot of an experiment independent of that shown in A. Corresponding DSB signals at YCR047c were quantified and superimposed to compare intensity and timing. The scale for DSBs was chosen so that the final level of DSBs corresponds to the final level of bound Spo11. Note the delayed increase of DSBs in the rad50S strain compared with cross-linking-independent association of Spo11-Myc to the hotspot region.