Figure 1
AML1 coexists with AML1/ETO on chromatin in t(8;21) leukemic cells. (A) Schematic diagrams of the antibody recognition sites to wild-type AML1 and the AML1/ETO fusion protein. The anti-AML1 (N20) antibody targets the N terminus of AML1 and recognizes both AML1 and AML1/ETO; the anti-AML1 (C19) antibody targets the C terminus of AML1 and recognizes AML1 but not AML1/ETO; and the anti-ETO (C20) antibody targets the C terminus of ETO and specifically recognizes AML1/ETO. The significant ChIP regions enriched by anti-ETO (C20), anti-AML1 (C19), and anti-AML1 (N20) antibody in Kausmi-1 cells are available in supplemental Tables 1 to 3, respectively. (B) Validation of antibodies specific to AML1, AML1/ETO, and both proteins by western blotting with the Kasumi-1 cell lysates. (C) Acquisition of high confidence AML1/ETO and AML1 binding sites in Kasumi-1 cells. High confidence AML1/ETO binding sites (supplemental Table 4) were generated by overlapping ChIP regions enriched by anti-ETO (C20) (supplemental Table 1) and anti-AML1 (N20) antibodies (supplemental Table 3) (upper). High confidence AML1 binding sites (supplemental Table 5) were generated by overlapping ChIP regions enriched by anti-AML (C19) (supplemental Table 2) and anti-AML1 (N20) antibodies (supplemental Table 3) (lower). (D) Venn diagram of the overlap between high confidence AML1/ETO and AML1 binding sites in Kasumi-1 cells. AML1/ETO∩AML1, overlapped ChIP regions. The full list of the overlapped high confidence ChIP regions between AML1/ETO and AML1 in Kasumi-1 cells are available in supplemental Table 6. (E) The genomic distribution of wild-type AML1 differs between AML1/ETO-positive and -negative cells. Each region bound by AML1 or AML1/ETO was mapped to the closest Refseq gene. upstream, upstream regulatory regions between −20 and −3 kb to the transcription start site (TSS); promoter, regions between −3 and 1 kb to the TSS; downstream, 20-kb regions downstream regulatory regions of the transcription termination site (TTS); exon and intron, regions mapped to related location according to Refseq annotations; intergenic, other regions. ChIP-seq data used in this analysis were retrieved from the NCBI Gene Expression Omnibus database (GSM610330,24GSM837994,23GSM722704,20 and GSM85082420).

AML1 coexists with AML1/ETO on chromatin in t(8;21) leukemic cells. (A) Schematic diagrams of the antibody recognition sites to wild-type AML1 and the AML1/ETO fusion protein. The anti-AML1 (N20) antibody targets the N terminus of AML1 and recognizes both AML1 and AML1/ETO; the anti-AML1 (C19) antibody targets the C terminus of AML1 and recognizes AML1 but not AML1/ETO; and the anti-ETO (C20) antibody targets the C terminus of ETO and specifically recognizes AML1/ETO. The significant ChIP regions enriched by anti-ETO (C20), anti-AML1 (C19), and anti-AML1 (N20) antibody in Kausmi-1 cells are available in supplemental Tables 1 to 3, respectively. (B) Validation of antibodies specific to AML1, AML1/ETO, and both proteins by western blotting with the Kasumi-1 cell lysates. (C) Acquisition of high confidence AML1/ETO and AML1 binding sites in Kasumi-1 cells. High confidence AML1/ETO binding sites (supplemental Table 4) were generated by overlapping ChIP regions enriched by anti-ETO (C20) (supplemental Table 1) and anti-AML1 (N20) antibodies (supplemental Table 3) (upper). High confidence AML1 binding sites (supplemental Table 5) were generated by overlapping ChIP regions enriched by anti-AML (C19) (supplemental Table 2) and anti-AML1 (N20) antibodies (supplemental Table 3) (lower). (D) Venn diagram of the overlap between high confidence AML1/ETO and AML1 binding sites in Kasumi-1 cells. AML1/ETO∩AML1, overlapped ChIP regions. The full list of the overlapped high confidence ChIP regions between AML1/ETO and AML1 in Kasumi-1 cells are available in supplemental Table 6. (E) The genomic distribution of wild-type AML1 differs between AML1/ETO-positive and -negative cells. Each region bound by AML1 or AML1/ETO was mapped to the closest Refseq gene. upstream, upstream regulatory regions between −20 and −3 kb to the transcription start site (TSS); promoter, regions between −3 and 1 kb to the TSS; downstream, 20-kb regions downstream regulatory regions of the transcription termination site (TTS); exon and intron, regions mapped to related location according to Refseq annotations; intergenic, other regions. ChIP-seq data used in this analysis were retrieved from the NCBI Gene Expression Omnibus database (GSM610330,24 GSM837994,23 GSM722704,20  and GSM85082420 ).

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