Fig. 2.
Interaction of NEIL2 with BER proteins. (A) Far-Western analysis. Proteins were separated by SDS-PAGE in duplicate gels, one of which was Coomassie stained. The proteins from the other gel were transferred to a nitrocellulose membrane and probed with NEIL2 as described in Section 2. (B) Yeast two-hybrid analysis of NEIL2 and Lig IIIα interaction. Yeast expressing NEIL2 and Lig IIIα fusion proteins (lanes 2 and 4) vs. control expressing only Lig IIIα fusion protein (lanes 1 and 3) were tested on medium with (lanes 1 and 2) and without (lanes 3 and 4) His. Other details are given in Section 2. (C) Right panel, in vitro pulldown of NEIL2 with His-tagged XRCC1 (lane 4) and His-tagged PNK (lane 5), respectively. Left panel, Western analysis using anti-His Ab showing the comparable amounts of His-tagged XRCC1 (lane 1) and PNK (lane 2). (D) Interaction of NEIL2 with full length and fragments of Pol β by Far-Western analysis. Pol β and its truncated fragments (10 µg) were separated by SDS-PAGE and Coomassie-stained (lane 1–6) and tested for interaction with NEIL2 (lanes 7–12). At right, a schematic showing the interacting Pol β domains.