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. 2007 Apr 15;21(8):898–903. doi: 10.1101/gad.1522607

Figure 3.

Figure 3.

Effect of modifying primer ends on Chk1 and Rad1 phosphorylation induced by primed M13 ssDNA. (A) Names and figures of structures used in B–D. Dot represents the biotinylated end of the primer. (B) M13, M13 + unmodified 80-mer, or M13 + 80-mer blocked at the 3′-end with biotin was incubated with streptavidin and added to NPE. Samples were taken and analyzed for phospho-Chk1 (S344), Chk1, or Rad1 by immunoblotting, or for replication as described in Figure 1A. (C) M13 ssDNA annealed to an 80-mer blocked at the 3′-end, 5′-end, or 3′- and 5′-ends with biotin were incubated with streptavidin, added to NPE in the presence of aphidicolin, and analyzed as described in B. (D) M13 ssDNA annealed to an 80-mer blocked at the 5′-end with biotin was incubated with streptavidin, added to NPE in the absence or presence of aphidicolin, and analyzed as described in B.