Figure 2

Interaction of N782 with the XRCC5/6 complex. * Indicates EGFP tag. (a) Left panel shows proteins immunoprecipitated with a rabbit anti-GFP antibody (black circle) and co-immunoprecipitated proteins (arrow). It contains both rabbit IgG heavy chain (50 kD wide band) and the light chain (25 kD wide band) derived from the anti-GFP antibody. Right panel shows Western blots of immunoprecipitated EGFP-KDM2A, EGFP-N782, and EGFP-KDM2A C-termini. The proteins were detected by a combination of mouse anti-GFP (primary) and goat anti-mouse IgG. (b) Keratinocyte genomic DNA fragments co-immunoprecipitated with EGFP-N782 and EGFP. A marker (250 or 500 ng) or sample (5 μl) was applied to each well, respectively. (c) Mass spectrometry of proteins included in the 70–90 kDa protein bands [proteins from top to bottom, (XRCC5 and XRCC6 in red) and (TMPO and LMNA in blue)]. (d) Western blotting analysis of XRCC5 (arrow) co-immunoprecipitated with N782 or the mutants (circle): Δ = deletion, C = CXXC, J = JmjC, and P = PHD. (e) Binding of GST-CN, GST-PN, and GST-NLSR to 3-length IGFBPL1 baits. Full-length blots are presented in Supplementary Fig. S1. (f) Far Western blotting analysis of the interaction of KMD2A domains with XRCC5 and 6. Probe and protein dye are on the left; cross-reaction of antibody to an E. coli protein, > and <. Full-length blots are presented in Supplementary Fig. S2. (g) Amino acid sequence of CPN domains expressed in cells (CPN, ⁵⁶¹V - R⁷³³; CN, ⁵⁶¹V- T⁶¹⁹ plus ⁶⁷⁶Y - R⁷³³; PN, ⁶¹⁹T - R⁷³³; NLSR, ⁶⁷⁶Y - R⁷³³). Typically defined domains C (red) and P (violet), and the N (blue). We have defined N by deletion analysis (unpublished).