Journal: Signal Transduction and Targeted Therapy
Article Title: Extraocular delivery of bioswitchable tri-miR-22-loaded tetrahedral DNA nanostructures for intraocular neovascular and neurodegenerative repair
doi: 10.1038/s41392-025-02566-4
Figure Lengend Snippet: Subconjunctival BiRDS suppressed retinal neovascularization and promoted healthy angiogenesis in the OIR mouse model. a Schematic description of the establishment of the OIR model and the design of the animal experiments. b Schematic representation of retinal flat-mount lesions (neovascularized and nonperfused areas) in OIR model mice and key morphological hallmarks of healthy angiogenesis (filopodia, tip cells, and stalk cells). c Upper: Retinal flat mounts after OIR and drug-treated OIR mice. Scale bar = 1 mm. Lower: higher-magnification images of pathological neovascular tufts. Scale bar = 50 μm. d Avascular area measured for quantification, as indicated by the dotted yellow lines. Scale bar = 1 mm. e Higher magnification images of pathological vessels sprouting from veins, as indicated by the dotted green line. Scale bar = 100 μm. f Upper: representative images of tip cells; the yellow arrowhead indicates tip cells. Scale bar = 50 μm. Lower: representative images of filopodia; yellow arrows indicate filopodia. Scale bar = 10 μm. g Retinal cryosections and immunofluorescence staining of drug-treated OIR mice. Scale bar = 50 μm. White: IB4 (vessels); red: CD31 (neovasculature); green: VEGFR; blue: DAPI (nuclei). h ‒ n Quantification of neovascular areas ( h ), avascular areas ( i ), sprouting areas ( j ), and tip cells ( k ) or counts of filopodia ( l ) and counts of neovascular cell nuclei anterior to the ILM ( m ) or vascular tube of the DCP ( n ). SCP, superficial capillary plexus; DCP, deep capillary plexus. Mean ± SD. n = 6. *** p < 0.001, ** p < 0.01
Article Snippet: The following antibodies were used for immunofluorescence: Isolectin GS-IB4, Alexa FluorTM 568( I21412 , Thermo Fisher Scientific, Massachusetts, USA), CD31(SC-376764, Santa Cruz Biotechnology, Dallas, TX, USA), VEGFR (#AF644; R&D Systems, Minneapolis, MN, USA), Tuj1 (#4466; Cell Signaling Technology, Danvers, MA, USA), GFAP (#12389; Cell Signaling Technology, Danvers, MA, USA), PKC-α (#sc-8393; Santa Cruz Biotechnology, Dallas, TX, USA), Rhodopsin (#ab221664; Abcam, Cambridge, UK), Calbindin (#ab82812; Abcam, Cambridge, UK), Alexa Fluor 488-labeled goat anti-rabbit IgG (#4412S; Cell Signaling Technology, Danvers, MA, USA), and Alexa Fluor 488-labeled goat anti-mouse IgG (#4408S; Cell Signaling Technology, Danvers, MA, USA), Alexa Fluor 555-conjugated goat anti-rabbit IgG (#25363S, Cell Signaling Technology, Danvers, MA, USA), Alexa Fluor 555-conjugated goat anti-mouse IgG (#37459S, Cell Signaling Technology, Danvers, MA, USA).
Techniques: Immunofluorescence, Staining
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