Journal: bioRxiv
Article Title: Heterochronic transcription factor expression drives cone-dominant retina development in 13-lined ground squirrels
doi: 10.1101/2025.04.25.650540
Figure Lengend Snippet: Mef2c is sufficient to promote cone-specific gene expression and repress rod-specific gene expression in mouse. ( A ) Diagram of overexpression strategy used to test effects of MEF2C overexpression. ( B ) UMAP representation of P8 mouse retinal explants electroporated with a plasmid expressing GFP alone (Empty) or GFP in a bicistronic transcript with human MEF2C ( MEF2C ), (n = 7445 cell Empty, 11949 cells MEF2C ). Each point represents a single cell and is colored by cell type as determined by clustering and marker gene expression. ( C ) Heatmap of expression for select genes for cones, cone-like photoreceptor precursors, rod photoreceptor precursors, and rods in cells overexpressing MEF2C , scaled by gene. ( D ) Immunohistochemistry showing GFP and GNAT2 expression in P8 mouse retinas from Empty and MEF2C conditions. Scale bars, 50 µm. ( E ) Box plot of the number of Gnat2+, GFP+ cells divided by the total number of GFP+ cells (n = 8 for both conditions). P-values calculated by Wilcoxon rank-sum test. P0, postnatal day 0; P8, postnatal day 8; FACS, fluorescence-activated cell sorting; scRNA-Seq, single-cell RNA sequencing; ONL, outer nuclear layer; INL, inner nuclear layer; DAPI, 4′,6-diamidino-2-phenylindole; GFP, green fluorescent protein; OE, overexpression.
Article Snippet: Primary antibodies utilized for staining 13LGS sections included goat anti-Otx2 polyclonal IgG (R&D Systems, BAF1979, 1:400) and rabbit anti-Mef2c polyclonal (Atlas Antibodies, HPA005533, 1:50).
Techniques: Gene Expression, Over Expression, Plasmid Preparation, Expressing, Marker, Immunohistochemistry, Fluorescence, FACS, RNA Sequencing
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