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. 2014 Jul 3;23(22):2700–2711. doi: 10.1089/scd.2014.0129

FIG. 6.

FIG. 6.

S-phase progression after the depletion of Rad51 protein. (A) mESCs transfected with siNS or siRad51 were incubated with 2 mM thymidine for 16 h and then released into fresh media. Two hours later, the thymidine analogs IdU (50 μM) and CldU (200 μM) were successively incorporated into the DNA replication sites. Subsequently, the colocalization of IdU and CldU replication foci was stained with corresponding antibodies and the degree of colocalizing foci areas was quantified in each isolated cell using Image J software (right). (B) mESCs transfected with siNS or siRad51 were successively pulse labeled with IdU and CldU to final concentrations of 100 and 250 μM, respectively, for 20 min. DNA fibers were immunostained with antibodies specific for IdU and CldU. The replication speed was quantified from the mean fork extension rate (kb/min) during sequential pulse labels with IdU (1st label, 20 min) and CldU (2nd label, 20 min) in mESCs.