HIF-1α binding in suspended MDA-MB-231 breast cancer cells

We report the transcriptional differences of suspended MCF-7 and MDA-MB-231 breast cancer cells after treated with CoCl2 100uM for 24 hours. CUT&Tag technology represents a cutting-edge method for investigating protein-DNA interactions. Utilizing a hyperactive novel fusion enzyme (Hyperactive pG-Tn5 / pA-Tn5 Transposase), this technique, under antibody guidance, specifically targets and cleaves DNA sequences near target proteins. It boasts several advantages over traditional ChIP-Seq, including lower cell input requirements, enhanced signal-to-noise ratio, and improved repeatability. MDA-MB-231 cells treated with 100uM CoCl2 for 24 hours were used. The CUT&Tag assay and subsequent DNA library construction were performed by following the manufacturer’s instructions for the Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903-01). An anti-HIF1-a monoclonal antibody (Affinity, BF8002) is used as the primary antibody. Paired-end sequencing was performed on an Illumina Novaseq 6000 system. Paired-end reads (150 bp) were aligned using Bowtie2 version 2.2.5. Macs2 peak calling software was used for peak calling.