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Integrated DNA Technologies alt-r crispr custom guide rnas
Alt R Crispr Custom Guide Rnas, supplied by Integrated DNA Technologies, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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transgenic constructs longgmr-cas9, customized guide rnas, and gmr-gcamp6s  (GenScript corporation)
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S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of <t>transgenic</t> inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.
Transgenic Constructs Longgmr Cas9, Customized Guide Rnas, And Gmr Gcamp6s, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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custom lipid metabolism–based guide rna library  (Twist Bioscience)
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S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of <t>transgenic</t> inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.
Custom Lipid Metabolism–Based Guide Rna Library, supplied by Twist Bioscience, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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custom single-guide rna (sgrna) lentiviral library  (Cellecta Inc)
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S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of <t>transgenic</t> inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.
Custom Single Guide Rna (Sgrna) Lentiviral Library, supplied by Cellecta Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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custom guide rna library  (GenScript corporation)
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S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of <t>transgenic</t> inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.
Custom Guide Rna Library, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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custom guide rnas (grnas  (Synthego Inc)
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S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of <t>transgenic</t> inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.
Custom Guide Rnas (Grnas, supplied by Synthego Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of transgenic inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.

Journal: bioRxiv

Article Title: Retinal Calcium Waves Coordinate Uniform Tissue Patterning of the Drosophila Eye

doi: 10.1101/2025.06.02.657513

Figure Lengend Snippet: S4A. Representative immunostaining images of Inx2 and Inx3 in 40hAPF retinas. These Z-stack images display the apical layer (A1–A6) , photoreceptor (PR) layer (A1’–A6’) , and basal layer (A1’’–A6’’) . Gap junction plaques containing Inx2 and Inx3 are observed at retinal support cell junctions, labeled with either Dlg1 (red) or E-cadherin (cyan). No Inx2 or Inx3 plaques were detected around PR cell bodies, indicated by empty space in the center of the ommatidium (A6’) , confirming that gap junctions are primarily in retinal non-neuronal cells. Scale bar: 10 μm (A1-6). S4B. Representative immunostaining images of Inx4–8 (Inx-X) in 40hAPF retinas, with innexins (white for B1’–5’) and adherens junctional marker E-cadherin (magenta for B1–4) or septate junctional marker Coracle (B5) . No significant Inx 4–8 signals were detected at ommatidial junctions at 40hAPF retina, with weak Inx5 signal from the early rhabdomere (B2) . Scale bar: 10 μm (B1-5). S4C. Immunostaining of Inx8 in retinas (C1) and photoreceptor axonal terminals (C2, C2’) at 48hAPF, using Inx8 ( green ) and neuronal membrane marker HRP ( blue ) , respectively. No meaningful signal was detected in the retina (C1) , suggesting the subcellular localization of Inx8 in PRs is confined to axonal terminals located in the optic lobe (C2) . Scale bar: 10 μm (C1,2). S4D. Representative images of retinal wave activity, displayed as color-coded 3-minute temporal calcium activity maps from control (D1) and inx8 mutant retinas (D2) during stage 2 waves. Scale bar: 10 μm (D1,2). See also Supplementary Movie S11 . S4E. Quantification of retinal calcium levels from S4D. n ≥ 6 retinas, ns: p ≥ 0.5. Error bars represent SD. S4F. Representative retinal images of transgenic inx1 mutants at 40hAPF, including control (F1, F1’), inx1-RNAi (F2, F2’), gRNA-inx1 (BDSC version) with longGMR-Cas9 (F3, F3’), gRNA-inx1 (custom version) with longGMR-Cas9 (referred as inx1 -sKO ) (F4, F4’), and gRNA-inx1 (BDSC version) with longGMR-G4 > UAS-Cas9 (F5, F5’) , using Inx1 (green) and junctional marker Dlg1 ( magenta ) , respectively. The images are Z-stacks of the retinal apical layer. Scale bar: 10 μm (F1-5). S4G–I. Quantification of Inx1 (G), Inx2 (H), and Inx3 (I) immunostaining signals in inx s-sKO mutant retinas from . n ≥ 14 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD. S4J. Quantification of vertical pathway frequency for each step indicated above in inx mutant retinas during late stage 1. All values were normalized to control. n ≥ 200 ommatidia from 5 retinas, ns: 0.5 ≥ p, *: p < 0.5, **: p < 0.01, ***: p < 0.001. Error bars represent SD.

Article Snippet: Generation of Transgenic Animals: Transgenic constructs including longGMR-Cas9, customized guide RNAs, and GMR-GCaMP6s were generated by GenScript (Piscataway, NJ, USA).

Techniques: Immunostaining, Labeling, Marker, Membrane, Activity Assay, Control, Mutagenesis, Transgenic Assay