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. 2018 Oct 26;1(5):e201800143. doi: 10.26508/lsa.201800143

Figure S5. Topological loading onto DNA without added ATP.

Figure S5.

(A) A DNA release experiment by restriction enzyme cleavage was performed following a cohesin loading reaction in the presence or absence of added ATP. The immobilized cohesin-DNA complex was incubated with or without PstI. Supernatant (S) and bead-bound (B) fractions were collected and DNA in each fraction was analyzed by agarose gel electrophoresis. (B) Gel image and quantification of the recovered DNA from cohesin loading reactions performed with or without added ATP using Scc2C as the cohesin loader. (C) Topological DNA loading by Walker B motif mutant EQ-cohesin, in the absence or presence of the cohesin loader, was analyzed by DNA release following linearization with PstI. Supernatant (S) and bead-bound fractions (B) were retrieved following PstI or mock treatment. DNA in both fractions was analyzed by agarose gel electrophoresis.