Fig 1: Discovery of a Novel SALL4 DNA-Binding Motif(A–C) DNA sequence motifs bound by WT SALL4A (A), a mutant lacking the 2nd zinc finger cluster (A?ZFC2) (B), or a mutant lacking the 4th ZFC (A?ZFC4) (C), discovered in universal PBM assays. The color bars above the position weighted matrices indicate the linear structure of SALL4, and the ZFCs are denoted by black ovals.(D) EMSA showing SALL4A shifts the AT-rich motif-containing oligos (lanes 1–3) but not when the motif is scrambled (lanes 4–6, gel cut for clarity); UC, unlabeled competitor probes.(E) SALL4-DNA complex is super-shifted by a SALL4 monoclonal antibody (lane 4) but not by mouse IgG isotype control (lane 5); mAb, SALL4 mouse antibody (Santa Cruz EE-30).(F) EMSA showing that SALL4-DNA complex was reduced or super-shifted in the presence of FLAG or SALL4 antibodies; rAb-1, SALL4 rabbit antibody (Cell Signaling D16H12); rAb-2, SALL4 rabbit antibody (Abcam ab57577). Lanes 8–10 show that the A?ZFC3 mutant binds to the same WT sequence, whereas binding by A?ZFC2 and A?ZFC4 mutants is abrogated. All EMSA reactions contain poly dI:dC competitor to reduce background binding.(G) Isothermal titration calorimetry (ITC) experiments showing purified SALL4 ZFC4 (amino acids 864–929) binds DNA oligos containing the WT WTATB motif (top) and not when the motif was mutated (bottom). All EMSA and ITC oligo sequences can be found in the Key Resources Table.
Supplier Page from Abcam for Anti-Sall4 antibody