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. 2015 Apr 28;34(13):1829–1843. doi: 10.15252/embj.201489865

Figure 6.

Figure 6

A FEN1/MutSα functional complex in the AEE assay
  1. RPR assays with the purified recombinant proteins of Pol δ, PCNA, RFC, FEN1, the MutSα complex, Ligase 1 (Lig I), and the substrates A1 and A2. 100 fmol of hPol δ, 300 fmol of hPCNA, 300 fmol of hRFC, 150 fmol of hMutSα, and 240 fmol of Lig I were mixed with 500 fmol of hFEN1 as indicated. The mixture was then incubated with 50 fmol of substrate in the reaction buffer containing 5 µCi [α-32P] dTTP and 50 µM each of the other three nucleotides. The reactions were carried out at 37°C for 60 min.
  2. AEE assays were identical to (A) except for the use of 5 µCi [α-32P] dGTP instead of 5 µCi [α-32P] dTTP.
Data information: Numbers at the bottom of the panels are the lane numbers for the various reactions. AV, the FEN1 mutant A159V. ED, the FEN1 mutant E160D. Lig. Prod., ligated product. Source data are available online for this figure.