Figure 5.

Immunodepletion of Pol δ from purified human whole cell extract fractions containing activity directed against 5-OHU². Active fraction (fraction 22) from Mono-Q chromatography containing 5-OHU² activity was mock-immunodepleted and immunodepleted using Pol δ specific antibodies and samples analysed by SDS-polyacrylamide gel electrophoresis and western blotting using antibodies against XPF and Pol δ. (A) The original fraction, mock-immunodepleted and Pol δ immunodepleted fraction was tested for 5-OHU² activity using 300 fmol of duplex oligonucleotide (B) Samples were incubated for 20 min at 37°C prior to the addition of formamide loading dye and analysis by 20% denaturing polyacrylamide gel electrophoresis and phosphorimaging.