MCF7 cells were cultured in normal or glucose-free media for 24 hours, and 6 × 106 cells were subjected to the CUT&RUN assay with anti-CTCF antibodies (Cell Signaling Technology, 3418) (55). Genomic DNA fragments (200 to 600 bp) were recovered from agarose gels. Library preparation and high-throughput sequencing were conducted by the Genomic Services Laboratory at HudsonAlpha. In replicate experiments, libraries were prepared using Illumina’s TruSeq ChIP Sample Preparation Kit (IP-202-1012) according to the manufacturer’s instructions. The quality of the library was checked with Agilent TapeStation. Final libraries were subjected to paired-end sequencing of 100-bp length on an Illumina HiSeq 2500 (30 to 40 million reads for each sample). The obtained genome-wide CTCF-binding data were deposited in the National Center for Biotechnology Information, NIH (NCBI) Gene Expression Omnibus (GEO; GSE114319).

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