Fig. 5

Genomic-wide analysis of Hey2-mediated chromatin occupancy, histone deacetylation and chromatin condensation in NRVMs.a Schematic diagram for CUT&Tag-seq and ATAC-seq analysis using Adv-Hey2-transduced or siRNAs-transfected NRVMs. b, c Heatmaps of H3K9ac and H3K27ac CUT&Tag-seq signals in Adv-Hey2-infected NRVMs compared to controls, using a 3?kb window centered on peak regions and sorted in descending order by signal intensity. d Venn diagram outlining 1484 overlapped genes between genes with HEY2 binding peaks and reduced H3K9 acetylation and/or H3K27 acetylation in the promoter regions. e, f The Genome browser views at locus of Ppargc1b and Esrra showing marks of Flag-HEY2, H3K9ac, H3K27ac CUT&Tag-seq and RNA-seq from Adv-Ctrl- and Adv-Hey2-infected NRVMs; and also showing marks of HDAC1 CUT&Tag, ATAC-seq from Ctrl-siRNA- and Hey2-siRNA (knockdown, KD)-transfected NRVMs or Adv-Ctrl- and Adv-Hey2-infected NRVMs. 2 biological replicates. g, h Hey2 or Hdac1 immunoprecipitation with anti-Flag antibody or anti-HA antibody, respectively. Hey2 or Hdac1 was detected using anti-Flag or anti-HA antibody. 3 independent experiments. i Co-immunoprecipitation analysis of Flag-HEY2 and endogenous HDAC1 in Adv-Ctrl- and Adv-Hey2-infected NRVMs. 3 independent experiments. j Luciferase activities in 293?T cells that were transfected with pGL3-PGC1A-Fluc, pGL3-ESRRA-Fluc or pGL3-CPT2-Fluc with control pGL3, pGL3-HEY2, pGL3-HDAC1, and pGL3-HEY2 and pGL3-HDAC1 together. The relative flyfire luciferase activity was normalized by Renilla luciferase and calculated as the ratio of each experimental group to the control group. 3 biological replicates. Data presents as mean?±?SD, *P?