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. 2005 Apr 14;33(7):2181–2191. doi: 10.1093/nar/gki513

Figure 4.

Figure 4

Analysis of NER activities of human CFE2s for Oxa and Oxa–Sp. (A) 150OXAe, 150OXA–SPe and 150FL (all 2 nM and 5′-end labeled, Figure 1C) were incubated with HeLa CFE2s (100 μg) at 30°C for 0, 20, 60 and 90 min, and products were analyzed by 10% denaturing PAGE. Note that the only gel region containing incised products is shown. The sequence of the 59mer marker is given in Figure 1C. (B) The amount of 5′ incision products shown in panel A was quantitated for Oxa–Sp (circles), FL (triangles) and Oxa (squares), and are plotted against incubation time. (C) 150OXA–SPi (2 nM and internally labeled, Figure 1C) was incubated with HeLa CFE2s (100 μg), and products were analyzed as in panel A. The sequences of a 64mer marker and a 27mer bearing Oxa–Sp are given in Figure 1C. The fragments derived from damage-specific 3′ and dual incisions are indicated by asterisks. The fragments from unknown origin are indicated by dots (see also text). (D) 150OXA–SPe (2 nM) was incubated with CFE2s (100 μg) from HeLa, XPA and XPF cells, or a mixture of CFE2s from XPA (50 μg) and XPF (50 μg) cells at 30°C for 90 min. Products were analyzed as in panel A. The CFE2s used are indicated on the gel.