Journal: bioRxiv
Article Title: Heart tube morphogenesis is regulated by segment-specific gene regulatory networks controlled by MEF2C
doi: 10.1101/2024.11.01.621613
Figure Lengend Snippet: A) Schematic of selection process to identify candidate MEFC-dependent enhancers from integrated snRNA-seq and snATAC-seq data. B) Genome browser tracks displaying snATAC-seq accessibility profiles, MEF2C ChIP-seq occupancy profiles , and H3K27ac ChIP-seq occupancy profiles at example loci containing IFT-specific (left), V-specific (middle), and OFT-specific (right) candidate enhancers (yellow highlights). C) Genome browser tracks displaying snATAC-seq accessibility profiles, MEF2C ChIP-seq occupancy profiles , and H3K27ac ChIP-seq occupancy profiles at the Myh6 / Myh7 locus, which contains two candidate enhancers (yellow highlights, MVEB1 and MVEB2) with IFT-specific altered accessibility that overlap with regions found in the VISTA Enhancer Browser database . D) Images of E11.5 mouse embryos from the Vista Enhancer Browser database demonstrating positive enhancer activity of regions that overlap with candidates MVEB1 and MVEB2. E) Schematic of the Tol2 Transgenesis assay used to screen candidate enhancers in zebrafish. Elements of this panel were created in BioRender. B, B. (2024) https://BioRender.com/h83r503 . F) Representative ventral view images of zebrafish embryos at 72 hpf injected with candidate enhancers that demonstrated positive activity in the heart. Boxed area in the representative brightfield image (left) indicates the anatomical region of interest captured in the fluorescent images. G) Schematic representation of the observed onset of enhancer activity for candidate enhancers that demonstrated positive activity in the heart. H) Representative ventral view images of zebrafish embryos at 24 and 72 hpf injected with MVEB2:eGFP or MVEB6:eGFP reporter constructs. Scale bars = 100 μm.
Article Snippet: The resulting fragments were cloned into the Tol2 transgenic vector E1b-eGFP-Tol2 ( ) (RRID: Addgene_37845) using the In-Fusion Cloning system (Takara Bio).
Techniques: Selection, ChIP-sequencing, Activity Assay, Injection, Construct
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