Supplementary Figure 6: The search for binding locus of Bach factors. | Nature Immunology

Supplementary Figure 6: The search for binding locus of Bach factors.

From: The transcription repressors Bach2 and Bach1 promote B cell development by repressing the myeloid program

Supplementary Figure 6

a. Schematic representation of the MARE-like sequences of Cebpb, Ahr, Ly96, Spic, Bach1, Cish and Spi1.The arrows denote MARE-like sequences. b. Specificity of anti-Bach1 monoclonal antibody. Monoclonal antibody N9648 against mouse Bach1 was generated by immunizing mice with recombinant mouse Bach1 fragment (amino acid residues 318-392) and by establishing hybridoma clones from the spleens. The specificity of the antibody was confirmed by western blotting of extracts prepared from wild-type and Bach1-deficient embryonic fibroblasts. c. EMSA showing binding of Bach1 to the putative MAREs. EMSA was carried out using total 100 ng proteins (MBP-Bach1 or MBP-Mafk) with oligonucleotide probe which contained the MARE-like sequences of Ahr or Cebpb. Arrows indicate bindings of Bach1 (homodimer), Bach1-MafK (heterodimer) and MafK (homodimer). WT and Mut indicate competitor DNAs with wild-type and mutated sequences, respectively.

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