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. Author manuscript; available in PMC: 2011 Feb 4.
Published in final edited form as: DNA Repair (Amst). 2009 Dec 16;9(2):109. doi: 10.1016/j.dnarep.2009.11.002

Fig. 5.

Fig. 5

(A) Immunoblotting for UVDE and known LP BER proteins. Cell extracts were prepared from human XPA-vector (lane 1) and XPA-UVDE (lane 2) cells and immunoblotted with antibody for N. crassa UVDE, Pol β, FEN1, XRCC1, PARP-1, LIG I, LIG III, and G3PDH as a loading control as described in Section 2. (B) Nicking activity of human XPA-vector and XPA-UVDE cell extract on CPD-containing ODN substrate. The in vitro cell extract-based nicking analysis was performed as described in Section 2. The incubation was conducted for 15 (lanes 3 and 6), 30 (lanes 4 and 7), and 60 min (lanes 5 and 8). The substrate (42-mer) and nicked product (14-mer) are shown. (C) Repair activity of human XPA-vector and XPA-UVDE cell extracts on the nicked CPD ODN substrate. The incubation was conducted for 5 (lanes 3 and 6), 15 (lanes 4 and 7), and 30 min (lanes 5 and 8). The position of repair intermediates and ligated product were shown. The mobility of the 42-mer marker was slightly faster than the ligated product due to the presence of 5′-phosphate. (D) Neutralization of Pol β in the in vitro cell extract-based LP BER. The in vitro repair reaction was conducted with pre-immune IgG (lanes 3–5) and anti-Pol β IgG (lanes 6–8) as described in Section 2. The incubation was conducted for 5 (lanes 3 and 6), 15 (lanes 4 and 7), and 30 min (lanes 5 and 8). The position of repair intermediates and ligated products are shown.