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. 2014 Mar 12;42(9):5657–5670. doi: 10.1093/nar/gku198

Figure 1.

Figure 1.

Enhancement of ID2 monoubiquitination by DNA. (A) FANCI or the ID2 complex was used as substrate in ubiquitination reactions. ϕX RFI DNA (45 nM) was added to the reaction with the ID2 complex in lanes 5 (3-h reaction) and 6 (18-h reaction). Fluorescent staining of proteins is shown on the top, and two different exposures of a western blot with α-HA are shown underneath. (B) ID2 complexes containing wild-type proteins, FANCD2 (K561R) or FANCI (K523R) were tested. (C) ID2 ubiquitination was examined in the presence of wild-type or mutant FANCL or without FANCL. (D) ID2 ubiquitination was tested with various forms of ϕX DNA (RF-replicative form; HIII-HaeIII digested RF DNA). For (B)–(D), fluorescent-stained images on top are from 18-h reactions, whereas the western blots show products from 3-h reactions.