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CUT&RUN LoV-U time-course profiling of Wnt transcriptional complex in ΔCTNNB1 HEK293T cells compared to wild-type

ABSTRACT: Wnt/β-catenin signaling is commonly understood to activate transcription through pre-accessible regulatory elements. To investigate whether this pathway also directly modulates chromatin states, we performed CUT&RUN in human embryonic kidney 293T (HEK293T) and matched ΔCTNNB1 cells to profile β-catenin-associated chromatin complexes following pathway activation. Cells were pre-treated with the PORCN inhibitor LGK974 (10 nM) for 24 hours and subsequently stimulated with the GSK3 inhibitor CHIR99021 (10 µM) for 30 minutes, 90 minutes, or 4 hours to activate Wnt/β-catenin signaling. Chromatin binding of β-catenin, pan-TCF/LEF (LEF1, TCF7, TCF7L1, and TCF7L2), CBP/p300, HDAC1, H3K27ac, and H3K4me1 was assessed across conditions. An IgG control was included for all experiments. For perturbation experiments, cells were additionally treated with either the CBP/p300 inhibitor A-485 (5 µM) or the HDAC inhibitor sodium butyrate (1 mM). For the rescue experiment, LGK treated ΔCTNNB1 cells were transfected with pcDNA3-S33Yβ-catenin (Addgene#19286) or the empty vector control for 24 hours. The dataset captures signal-dependent and signal-independent changes in factor occupancy at regulatory regions, including conventional Wnt-responsive elements; and latent enhancers where β-catenin functions as a pioneer-like factor together with HDAC1 and CBP, mediating chromatin remodeling and TCF/LEF occupancy.

INSTRUMENT(S): KAPA EvoPrep Kit, Illumina HiSeq 2000

ORGANISM(S): Homo sapiens

SUBMITTER: Tamina Weiss

PROVIDER: E-MTAB-17186 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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