CUT&RUN was performed as described above. Peaks were called with MACS2. Heatmaps show CTCF peaks relative to IgG negative control antibody in aligned rows ranked by intensity (top to bottom) and colored such that red indicates high localized enrichment and blue denotes background signal. CTCF showed expected enrichment around the TSS. Despite some observable background in the IgG control, differential enrichment with the target antibody is as expected.
CUT&RUN was performed as described above. Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). Three representative loci are shown.
Figure 3: Immunohistochemistry data
FFPE sections of human colon carcinoma (left) and mouse spleen (right) using CTCF antibody at a dilution of 1:1000.
Figure 4: Immunocytochemistry data
FFPE section of human MCF-7 cells examined using CTCF antibody at a dilution of 1:1,000.
Figure 5: Immunoprecipitation data
EpiCypher CTCF antibody (6 μg/mg lysate) was used to immunoprecipitate whole cell lysates (1 mg, 20% of IP loaded) isolated from HEK293T cells. A negative control IgG antibody and positive control antibodies (Bethyl Laboratories) were also used to demonstrate specificity of the IP. All CTCF antibodies target the same CTCF epitope, however, A300-534A-6 is hosted in rabbit. For blotting immunoprecipitates, EpiCypher CTCF antibody was used at a 1:1,000 dilution.
Figure 6: Western blot data
Western analysis of CTCF in whole cell extracts from HEK293T, MCF-7, Jurkat, Hep-G2, and A-549 cells. Fifty micrograms of lysate was resolved via SDS-PAGE and detected with a 1:1,000 dilution of CTCF antibody.
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