Fig. 5.

Drosha is required for DNA resection after DNA damage and is recruited to DSBs. a Cartoon describing quantitative DNA resection assay. Treatment of U2OS cells with 4-OHT induces damage by HA-ER-AsiSI. Genomic DNA is harvested and subjected to restriction digest (dotted lines). qPCR primers, red lines, are designed at either side of the restriction sites. Only loci that had been resected prior to digest can be amplified. b qPCR resection assay as in a at two HR sites, and the control locus (no DSB) which does not span an AsiSI restriction site, as described in ref. ⁷ following 4 h of damage induction. Error bars = SD, Student’s 2-sample T-test, *p ≤ 0.05. c qPCR of Drosha and control IgG ChIP at an HR locus and a canonical Drosha binding site at the miR-122 genomic locus 1 h after damage induction. The ChIP efficiency was calculated against a histone H3 ChIP performed in parallel, error bars = SEM, Student’s 2-sample T-test, **p ≤ 0.01, N = 3