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. 2017 Jan 30;114(7):E1062–E1071. doi: 10.1073/pnas.1617309114

Fig. 5.

Fig. 5.

Transcriptional induction of URA3 abolishes cohesion but not cohesin binding. (A) Cohesion of DNA circles bearing the URA3 gene. Strains MSB7 (URA3f) and MSB20 (URA3r) were grown in media containing or lacking uracil from the outset of the experiment. (B) ChIP of Mcd1-TAP at chromosomal URA3 genes. Strain MSB37 (URA3e) was grown in media containing or lacking uracil from the outset and cross-linked with formaldehyde following M phase arrest by Cdc20 depletion. (C and D) Strains MSB19 (URA3f) and JSC230 (URA3r) were evaluated as in B. (E) Transcription-dependent loss of cohesin from URA3 in the absence of Scc2. Strain MSB128 (SCC2-AID-9myc ADH1p-OsTIR1) was arrested in M phase by Cdc20 depletion. Three hours later the culture was split into thirds. DMSO was added to one third, and 500 μM NAA was added to another. Uracil was removed from the final third when 500 μM NAA was added. Cohesin levels were assessed 2 h later at the 5′ end of the endogenous URA3 gene, as well as at a site on chromosome III (CARC1) that binds cohesin stably (42).