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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and <t>tuj1+</t> RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01
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Subconjunctival BiRDS rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and tuj1+ RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** p < 0.01

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 rescued retinal neurons in the OIR mouse model. a Schematic diagram of retinal neuron detection via immunofluorescence after subconjunctival injection of BiRDs. b Retinas from OIR mice were visualized through immunofluorescence staining at P17. Representative images of GFAP+ glial cells (red) and tuj1+ RGCs (green) in retinal sections (top), PKC-α+ rod bipolar cells (green) and rhodopsin+ rod cells (red) (middle), and calbindin+ horizontal cells (green) (bottom). DAPI (blue) was used to stain the retinal layers. c – g Semiquantification of immunofluorescence intensity in glial cells ( c ), RGCs ( d ), rod bipolar cells ( e ), rod photoreceptor cells ( f ), and horizontal cells ( g ). Scale bars = 50 μm. Mean ± SD. n = 6. **** p < 0.0001, ** 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, Injection, Staining