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. 2014 Sep 16;111(39):14118–14123. doi: 10.1073/pnas.1321349111

Fig. 4.

Fig. 4.

Nick translation by Pol δ and FEN1 in the presence of heterotrimeric PCNA. (A) Schematic illustration of the assay. PCNA was loaded on a radioactively labeled oligonucleotide substrate by RFC; Pol δ and FEN1 were added and the reaction was allowed to proceed for the indicated times. Pol δ elongates the primer until reaching the blocking oligo, generating a 54-nucleotide product. Then, strand displacement synthesis through the blocking oligo proceeds with or without FEN1, generating a final product of 84 nucleotides. Yellow star denotes radioactive label at 5′ end of the primer. The red segment represents the RNA portion of the blocking oligo. (B) Results of nick translation assay in the absence of PCNA or in the presence of four different PCNA trimer species. Pol δ and FEN1 were added where indicated. Reactions were analyzed by urea-PAGE and autoradiography.