FIGURE 7. xStn1 immunodepletion does not affect DNA polymerase α-primase binding to ssDNA in Xenopus egg extracts. A, M13 was incubated for 1 h in xStn1-immunodepleted or mock-immunodepleted HSS supplemented with 0.2 mm aphidicolin. The reactions were terminated by transferring the test tubes onto ice, and samples were immediately fractionated by gel filtration. Fractions were collected, and the presence of DNA was revealed by agarose gel electrophoresis followed by SYBR Gold staining. Identities of the circular/linearized forms were determined on the basis of the results shown in supplemental Figs. S4B and S7A. B, fractions 12–14 in A were analyzed by immunoblotting using the indicated antibodies. HSS that did not contain M13 DNA was also processed as described in A and used for the immunoblotting experiments (lanes 7-9). As a reference, 0.2 μl of HSS was loaded in lane 10.
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