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Proteintech rs1 polyclonal antibody
Rs1 Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rs1+polyclonal+antibody/RS1+Antibody/pm38661546-61-1-7
Average 93 stars, based on 5 article reviews

rs1 polyclonal antibody - by Bioz Stars, 2026-09

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( A ) RS1 expression variability and schisis cavity closure in XLRS mouse retina under mini-mGluR6 promoter . RS1 expression in WT and XLRS mouse retinas 5 weeks after intravitreal injection of AAV2/2-Y444F vector encoding human RS1 gene under bipolar cell specific mini-mGluR6 (In4s-In3-200En-mGluR500P) promoter. Retinal sections from injected XLRS mice along with age matched C57BL/6 WT mice were immunolabeled with antibodies raised against RS1 ( red ), Na/K ATPase a3 subunit ( green ), and DAPI counterstain for nuclei ( blue ). In WT retina, Rs1 is profusely present in the photoreceptor ISs, outer plexiform layer (OPL), bipolar cell layer, and IPL. In XLRS mouse retinas, intravitreal injection of viral vector leads to RS1expression specifically in inner retina with Rs1 immunoreactivity in cell processes within the IPL with moderate to modest cavity closure. RS1 localization is also seen in IS of photoreceptors. Retina 3 has the least cavity closure even though it has the most RS1 labeling. ( B) RS1 expression in transgenic mice under mini mGluR6 promoter. In transgenic mice the mini-mGluR6 promoter drives RS1 gene expression in the inner retina but the transgenic mice tended to have significantly more variability in gene expression levels with no effect on cavity closure. RS1 labeling is also seen in photoreceptor IS. Creation of transgenic mice is described in Materials and Methods section. ( C) RS1 localization on photoreceptor IS. Mini-mGluR6-RS1 injected XLRS mice retinas showed immunolabeling of Na/K ATPase ( green ) in photoreceptor IS, OPL and IPL, the labeling being intense in IPL. Merged images confirmed a high degree of colocalization of the Na/K ATPase ( green ) and RS1 ( red ) in all layers of the retina.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Expression of Retinoschisin by Retinal Bipolar Cells in XLRS Promotes Resolution of Retinoschisis Cysts Sans RS1 From Photoreceptors

doi: 10.1167/iovs.63.11.8

Figure Lengend Snippet: ( A ) RS1 expression variability and schisis cavity closure in XLRS mouse retina under mini-mGluR6 promoter . RS1 expression in WT and XLRS mouse retinas 5 weeks after intravitreal injection of AAV2/2-Y444F vector encoding human RS1 gene under bipolar cell specific mini-mGluR6 (In4s-In3-200En-mGluR500P) promoter. Retinal sections from injected XLRS mice along with age matched C57BL/6 WT mice were immunolabeled with antibodies raised against RS1 ( red ), Na/K ATPase a3 subunit ( green ), and DAPI counterstain for nuclei ( blue ). In WT retina, Rs1 is profusely present in the photoreceptor ISs, outer plexiform layer (OPL), bipolar cell layer, and IPL. In XLRS mouse retinas, intravitreal injection of viral vector leads to RS1expression specifically in inner retina with Rs1 immunoreactivity in cell processes within the IPL with moderate to modest cavity closure. RS1 localization is also seen in IS of photoreceptors. Retina 3 has the least cavity closure even though it has the most RS1 labeling. ( B) RS1 expression in transgenic mice under mini mGluR6 promoter. In transgenic mice the mini-mGluR6 promoter drives RS1 gene expression in the inner retina but the transgenic mice tended to have significantly more variability in gene expression levels with no effect on cavity closure. RS1 labeling is also seen in photoreceptor IS. Creation of transgenic mice is described in Materials and Methods section. ( C) RS1 localization on photoreceptor IS. Mini-mGluR6-RS1 injected XLRS mice retinas showed immunolabeling of Na/K ATPase ( green ) in photoreceptor IS, OPL and IPL, the labeling being intense in IPL. Merged images confirmed a high degree of colocalization of the Na/K ATPase ( green ) and RS1 ( red ) in all layers of the retina.

Article Snippet: The following antibodies were used: rabbit polyclonal antibody against the N-terminus of RS1 (amino acid residues 24–37 [1:1000; Custom made, ThermoFisher Scientific, Waltham, MA, USA] ; monoclonal antibody against the protein kinase C-alpha [1:500, Santa Cruz Biotechnology, Dallas, TX, USA]; mouse monoclonal antibody against glutamine synthetase [1:1000; Sigma-Aldrich, St. Louis, MO]; calretinin [1:2000; BD Biosciences, San Jose, CA, USA]; goat anti-mCherry [1:2000; Biorbyt, Cambridge, UK], and 1:1000 mouse monoclonal Na+/K+ ATPase [alpha-3 subunit] antibody [Na/K-ATPase alpha 3; ThermoFisher Scientific], secondary antibody conjugated to Alexa Fluor 568 dye or Alexa Fluor 488 dye [ThermoFisher Scientific]).

Techniques: Expressing, Injection, Plasmid Preparation, Immunolabeling, Labeling, Transgenic Assay, Gene Expression

MiniPromoter Ple155 drives bipolar cell specific RS1 gene expression in XLRS mice retinas . Retinal sections from representative vector injected and uninjected XLRS mice 5 weeks after subretinal injection of AAV8-Ple155-RS1 vector were immunolabeled with anti-RS1 antibody ( red ), RBC marker protein kinase Cα ( green ), Müller glia marker glutamine synthetase (GS), ( green ) and DAPI counterstain for nuclei ( blue ). Subretinal injection of AAV8-Ple155-RS1 at postnatal day 14, led to robust RS1 expression in bipolar cells ( a , e ) as confirmed by costaining with the bipolar cell marker PKCα ( b ). Compared with uninjected mice retina ( c , d ), which showed no RS1 labeling and displayed large schisis cavities, injected retinas showed schisis closure and marked improvement in inner retina structure. In AAV 8-Ple155-RS1 injected retinas, RS1 staining juxtaposed in close physical association with parallel Müller cell processes as revealed with anti-GS staining ( f , h ). Representative images for 4 mice in each group. The images in the lower panel show RS1 expression variability and photoreceptor localization in Ple155 injected retinas ( n = 4).

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Expression of Retinoschisin by Retinal Bipolar Cells in XLRS Promotes Resolution of Retinoschisis Cysts Sans RS1 From Photoreceptors

doi: 10.1167/iovs.63.11.8

Figure Lengend Snippet: MiniPromoter Ple155 drives bipolar cell specific RS1 gene expression in XLRS mice retinas . Retinal sections from representative vector injected and uninjected XLRS mice 5 weeks after subretinal injection of AAV8-Ple155-RS1 vector were immunolabeled with anti-RS1 antibody ( red ), RBC marker protein kinase Cα ( green ), Müller glia marker glutamine synthetase (GS), ( green ) and DAPI counterstain for nuclei ( blue ). Subretinal injection of AAV8-Ple155-RS1 at postnatal day 14, led to robust RS1 expression in bipolar cells ( a , e ) as confirmed by costaining with the bipolar cell marker PKCα ( b ). Compared with uninjected mice retina ( c , d ), which showed no RS1 labeling and displayed large schisis cavities, injected retinas showed schisis closure and marked improvement in inner retina structure. In AAV 8-Ple155-RS1 injected retinas, RS1 staining juxtaposed in close physical association with parallel Müller cell processes as revealed with anti-GS staining ( f , h ). Representative images for 4 mice in each group. The images in the lower panel show RS1 expression variability and photoreceptor localization in Ple155 injected retinas ( n = 4).

Techniques: Gene Expression, Plasmid Preparation, Injection, Immunolabeling, Marker, Expressing, Labeling, Staining

Functional and structural changes in XLRS mice retinas after injection of AAV8 vector encoding RS1 under Ple155 promoter . ( A ) Representative ERG waveforms recorded in scotopic and photopic conditions in 3 XLRS mice between 5 and 6 weeks after unilateral subretinal injection of AAV8-PLe155-RS1. The flash luminances at which the dark- and light-adapted ERGs shown in the figure were elicited were −0.82 log sc cd-s/m 2 and 1.0 log sc cd-s/m 2 , respectively. Both scotopic and photopic ERG responses in the injected eye ( red waveforms ) were increased in amplitude when compared with the uninjected eye ( black waveforms ). All three study animals showed larger scotopic and photopic b-wave amplitudes and a shorter scotopic a-wave implicit time in the treated eyes, indicating a selective improvement of the postphotoreceptorial element function, including the photoreceptor–bipolar cell synapse. The a-wave amplitude is measured from baseline (0 ms) to the negative trough; b-wave amplitude is measured from the a-wave trough to the positive peak. Stimulus flash occurs at 0 ms. ( B ) Analysis of retinal structure by OCT in the same 3 animals showed a reduction in intraretinal cystic cavities in the eye injected with the vector and almost complete restoration of the normal retinal architecture.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Expression of Retinoschisin by Retinal Bipolar Cells in XLRS Promotes Resolution of Retinoschisis Cysts Sans RS1 From Photoreceptors

doi: 10.1167/iovs.63.11.8

Figure Lengend Snippet: Functional and structural changes in XLRS mice retinas after injection of AAV8 vector encoding RS1 under Ple155 promoter . ( A ) Representative ERG waveforms recorded in scotopic and photopic conditions in 3 XLRS mice between 5 and 6 weeks after unilateral subretinal injection of AAV8-PLe155-RS1. The flash luminances at which the dark- and light-adapted ERGs shown in the figure were elicited were −0.82 log sc cd-s/m 2 and 1.0 log sc cd-s/m 2 , respectively. Both scotopic and photopic ERG responses in the injected eye ( red waveforms ) were increased in amplitude when compared with the uninjected eye ( black waveforms ). All three study animals showed larger scotopic and photopic b-wave amplitudes and a shorter scotopic a-wave implicit time in the treated eyes, indicating a selective improvement of the postphotoreceptorial element function, including the photoreceptor–bipolar cell synapse. The a-wave amplitude is measured from baseline (0 ms) to the negative trough; b-wave amplitude is measured from the a-wave trough to the positive peak. Stimulus flash occurs at 0 ms. ( B ) Analysis of retinal structure by OCT in the same 3 animals showed a reduction in intraretinal cystic cavities in the eye injected with the vector and almost complete restoration of the normal retinal architecture.

Techniques: Functional Assay, Injection, Plasmid Preparation

Human and mouse XLRS . ( A ) Retina photograph of a 17-year-old XLRS subject with 20/63 acuity showing classic macular retinoschisis with a subtle spoke wheel pattern radiating from the fovea. ( B ) Representative dark-adapted ERG combined responses (arising from photoreceptors and bipolar cells) of the XLRS subject show characteristic b-wave reduction disproportionate to a-wave reduction. ( C ) RS1 expression in mouse retina and XLRS phenotype. Retinal cryosections were immunolabeled with antibodies against RS1 ( red , 1:1000); Na/K ATPase a3 subunit ( green , 1:1000). The nuclei were counterstained with DAPI ( blue ). In WT retina RS1 is profusely expressed in photoreceptor IS and in inner retinal layers. In XLRS mouse retina, loss of Rs1 expression results in splitting of the inner retina cell layers. ( D ) RS1 expression on mouse retina bipolar cells and its colocalization with Na/K-ATPase, a plasma membrane marker. (E) Discoidin domain has been shown to be involved in cell adhesion during the streaming and aggregation of D. discoideum . During the growth phase of development, D. discoideum amoeboid cells feed on bacteria and replicate by binary fission. The development cycle is initiated upon starvation (resource depletion), and aggregation occurs when starving cells secrete cyclic AMP to recruit additional cells. Discoidin I is synthesized profusely as cells stream together into aggregate to form slug and fruiting body. OPL, outer plexiform layer; IPL, inner plexiform layer; GCL, ganglion cell layer. E is reproduced from Dunn JD, Bosmani C, Barisch C, et al. Eat prey, live: Dictyostelium discoideum as a model for cell-autonomous defenses. Front Immunol . 2018;8:1906. Copyright © 2018 Dunn, Bosmani, Barisch, Raykov, Lefrançois, Cardenal-Muñoz, López-Jiménez and Soldati; open-access, distributed under the terms of the Creative Commons Attribution License (CC BY).

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Expression of Retinoschisin by Retinal Bipolar Cells in XLRS Promotes Resolution of Retinoschisis Cysts Sans RS1 From Photoreceptors

doi: 10.1167/iovs.63.11.8

Figure Lengend Snippet: Human and mouse XLRS . ( A ) Retina photograph of a 17-year-old XLRS subject with 20/63 acuity showing classic macular retinoschisis with a subtle spoke wheel pattern radiating from the fovea. ( B ) Representative dark-adapted ERG combined responses (arising from photoreceptors and bipolar cells) of the XLRS subject show characteristic b-wave reduction disproportionate to a-wave reduction. ( C ) RS1 expression in mouse retina and XLRS phenotype. Retinal cryosections were immunolabeled with antibodies against RS1 ( red , 1:1000); Na/K ATPase a3 subunit ( green , 1:1000). The nuclei were counterstained with DAPI ( blue ). In WT retina RS1 is profusely expressed in photoreceptor IS and in inner retinal layers. In XLRS mouse retina, loss of Rs1 expression results in splitting of the inner retina cell layers. ( D ) RS1 expression on mouse retina bipolar cells and its colocalization with Na/K-ATPase, a plasma membrane marker. (E) Discoidin domain has been shown to be involved in cell adhesion during the streaming and aggregation of D. discoideum . During the growth phase of development, D. discoideum amoeboid cells feed on bacteria and replicate by binary fission. The development cycle is initiated upon starvation (resource depletion), and aggregation occurs when starving cells secrete cyclic AMP to recruit additional cells. Discoidin I is synthesized profusely as cells stream together into aggregate to form slug and fruiting body. OPL, outer plexiform layer; IPL, inner plexiform layer; GCL, ganglion cell layer. E is reproduced from Dunn JD, Bosmani C, Barisch C, et al. Eat prey, live: Dictyostelium discoideum as a model for cell-autonomous defenses. Front Immunol . 2018;8:1906. Copyright © 2018 Dunn, Bosmani, Barisch, Raykov, Lefrançois, Cardenal-Muñoz, López-Jiménez and Soldati; open-access, distributed under the terms of the Creative Commons Attribution License (CC BY).

Techniques: Expressing, Immunolabeling, Clinical Proteomics, Membrane, Marker, Bacteria, Synthesized

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Primers for Rs1 -KO genotyping.

Article Snippet: Proteins were detected by primary antibodies: polyclonal antibody against human RS1 (Atlas Antibodies, HPA059546, dilution 1:750) and β-Actin antibody monoclonal IgG sc-47778 (Santa Cruz Biotechnology, dilution 1:1000) then secondary antibodies: IRDye ® 800CW goat anti-rabbit IgG and 680RD goat anti-mouse IgG, dilutions 1:10000 (LI-COR).

Techniques: Sequencing

Expression cassette of the rAAV2tYF-CB-h RS1 vector. Components of rAAV2tYF-CB-h RS1 , the vector contains AAV serotype 2 inverted terminal repeats (ITRs) and an expression cassette consisting of a cytomegalovirus enhancer (CMV) and chicken β-actin promoter (CB), human RS1 complementary DNA, and a simian vacuolating virus 40 polyadenylation (SV40 PolyA) sequence and is packaged in an AAV2 capsid containing tyrosine-to-phenylalanine mutations in 3 surface-exposed tyrosine residues (Y444F, Y500F, Y730F) in the VP3 protein capsid (rAAV2tYF).

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Expression cassette of the rAAV2tYF-CB-h RS1 vector. Components of rAAV2tYF-CB-h RS1 , the vector contains AAV serotype 2 inverted terminal repeats (ITRs) and an expression cassette consisting of a cytomegalovirus enhancer (CMV) and chicken β-actin promoter (CB), human RS1 complementary DNA, and a simian vacuolating virus 40 polyadenylation (SV40 PolyA) sequence and is packaged in an AAV2 capsid containing tyrosine-to-phenylalanine mutations in 3 surface-exposed tyrosine residues (Y444F, Y500F, Y730F) in the VP3 protein capsid (rAAV2tYF).

Techniques: Expressing, Plasmid Preparation, Virus, Sequencing

Characterization of retinal phenotypes in retinoschisin-1 knockout mice: rod and cone photoreceptors functional loss and cyst formation. (A) OCT images of a Rs1- KO mouse (1) and a wild-type (WT) control littermate (2) are shown at 1 month of age. The outer nuclear layer (ONL) thicknesses are indicated by a white solid bar, and cysts are indicated by white stars on the OCT image. (B) Quantification of ONL thicknesses in Rs1 -KO and WT or heterozygous control mice at 1 month of age. Rs1 -KO mice have thinner ONLs. (C) Quantification of the cyst severity in Rs1 -KO and WT or heterozygous control mice. Cyst severity score is modified from Bush et al. where (1) no cavities; (2) <30 μm; (3) 30–49 μm; (4) 50–69 μm; (5) 70–99 μm; (6) ≥100 μm. Rs1 -KO mice have cysts whereas WT or heterozygous do not. (D) (1) Waveforms in dark-adapted (DA) standard combined response (SCR) ERG using 3.0 cd·s/m 2 stimuli in Rs1 -KO and WT or heterozygous control mice. (2) b/a wave ratio is reduced in Rs1 -KO compared to WT or heterozygous controls. (E) At 1 month of age, Rs1 -KO mice have greater a-wave amplitudes compared to WT or heterozygous control mice after 0.01 cd·s/m 2 stimuli in DA conditions. Representative waveforms are shown in (1), and the statistical comparison of their a-wave amplitudes is shown in (2). (F) Cone function on ERG measured after light adaptation using 3.0 flash (1) and 5 Hz flicker (2). Amplitudes are reduced in Rs1 -KO compared to WT or heterozygous control mice. MO: month old, Rs1 -KO: retinoschisin-1 knockout, WT: wild-type or heterozygous mice, μV: Microvolt, * p < 0.05, ** p < 0.01, and **** p < 0.0001. Scale bar, 100 μm.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Characterization of retinal phenotypes in retinoschisin-1 knockout mice: rod and cone photoreceptors functional loss and cyst formation. (A) OCT images of a Rs1- KO mouse (1) and a wild-type (WT) control littermate (2) are shown at 1 month of age. The outer nuclear layer (ONL) thicknesses are indicated by a white solid bar, and cysts are indicated by white stars on the OCT image. (B) Quantification of ONL thicknesses in Rs1 -KO and WT or heterozygous control mice at 1 month of age. Rs1 -KO mice have thinner ONLs. (C) Quantification of the cyst severity in Rs1 -KO and WT or heterozygous control mice. Cyst severity score is modified from Bush et al. where (1) no cavities; (2) <30 μm; (3) 30–49 μm; (4) 50–69 μm; (5) 70–99 μm; (6) ≥100 μm. Rs1 -KO mice have cysts whereas WT or heterozygous do not. (D) (1) Waveforms in dark-adapted (DA) standard combined response (SCR) ERG using 3.0 cd·s/m 2 stimuli in Rs1 -KO and WT or heterozygous control mice. (2) b/a wave ratio is reduced in Rs1 -KO compared to WT or heterozygous controls. (E) At 1 month of age, Rs1 -KO mice have greater a-wave amplitudes compared to WT or heterozygous control mice after 0.01 cd·s/m 2 stimuli in DA conditions. Representative waveforms are shown in (1), and the statistical comparison of their a-wave amplitudes is shown in (2). (F) Cone function on ERG measured after light adaptation using 3.0 flash (1) and 5 Hz flicker (2). Amplitudes are reduced in Rs1 -KO compared to WT or heterozygous control mice. MO: month old, Rs1 -KO: retinoschisin-1 knockout, WT: wild-type or heterozygous mice, μV: Microvolt, * p < 0.05, ** p < 0.01, and **** p < 0.0001. Scale bar, 100 μm.

Techniques: Knock-Out, Functional Assay, Control, Modification, Comparison

Expression of human RS1 protein in treated retinas of Rs1 -KO mice reveals a dose-dependent response. (A) Immunoblot analysis of eyes treated with 8E9 vg, 8E8 vg, 8E7 vg doses of the AAV2tYF vector, and diluent-treated samples (Dil) retinal protein extracts. Bleb qualities were excellent for all treatment groups except for one retina treated with 8E7 dose that had a very good bleb (arrowhead). Completely untreated Rs1 -KO and WT retinas served as negative and positive controls. Each lane represents an individual eye, with three eyes analyzed per dose, 2 eyes for sham treatment, 2 eyes for WT, and 1 eye for untreated KO. Eighteen μg of protein were loaded per lane. The expected molecular weights for proteins are indicated on the left, and the antibodies used to visualize the proteins are indicated on the right. (B) RS1 expression levels relative to completely WT eyes. Quantification involved normalizing the RS1 band intensity to the actin band intensity for each sample, with the expression level presented as a percentage of WT retinal samples which is set to 100%. The results illustrate a dose-dependent increase in RS1 protein expression in Rs1 -KO mouse retinas treated with 8E7, 8E8, and 8E9 vg of rAAV2-tYF-CB-h RS1 .

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Expression of human RS1 protein in treated retinas of Rs1 -KO mice reveals a dose-dependent response. (A) Immunoblot analysis of eyes treated with 8E9 vg, 8E8 vg, 8E7 vg doses of the AAV2tYF vector, and diluent-treated samples (Dil) retinal protein extracts. Bleb qualities were excellent for all treatment groups except for one retina treated with 8E7 dose that had a very good bleb (arrowhead). Completely untreated Rs1 -KO and WT retinas served as negative and positive controls. Each lane represents an individual eye, with three eyes analyzed per dose, 2 eyes for sham treatment, 2 eyes for WT, and 1 eye for untreated KO. Eighteen μg of protein were loaded per lane. The expected molecular weights for proteins are indicated on the left, and the antibodies used to visualize the proteins are indicated on the right. (B) RS1 expression levels relative to completely WT eyes. Quantification involved normalizing the RS1 band intensity to the actin band intensity for each sample, with the expression level presented as a percentage of WT retinal samples which is set to 100%. The results illustrate a dose-dependent increase in RS1 protein expression in Rs1 -KO mouse retinas treated with 8E7, 8E8, and 8E9 vg of rAAV2-tYF-CB-h RS1 .

Techniques: Expressing, Western Blot, Plasmid Preparation

rAAV2tYF subretinal gene therapy improves and sustains cone photoreceptor function in Rs1 -KO mice treated with the medium dose of 8E8 vg/eye over the course of 12 months. (A) Cone-dependent retinal function, assessed through light-adapted (LA) electroretinography after subjecting the eyes to a 3.0 cd·s/m 2 bright flash. Eyes treated with the 8E8 vg dose consistently displayed significantly greater b-wave amplitudes compared to sham-treated eyes and untreated eyes over 1 year. Eyes treated with 8E9 showed significance compared to the sham-treated eyes only at the experimental endpoint. (B) Representative LA waveforms were shown for eyes treated with different doses, as well as for sham-treated, and untreated eyes at 3 MPI and 7 MPI, respectively. (C) Another metric used to evaluate cone-dependent retinal function was the 5 Hz flicker test, which consistently showed that 8E8 vg dose-treated eyes outperformed sham-treated and untreated eyes throughout the treatment duration. Neither 8E9 nor 8E7 vg doses showed significance compared to the sham-treated eyes over the treatment course. (D) When subjected to a 5 Hz flicker, cones in treated eyes elicited higher amplitudes at 3 MPI, which was not observed in the sham-treated eyes or the untreated contralateral eyes. Over the course of 7 months, treated eyes retained significantly better amplitudes in cone ERGs. (E) Comparison of ERG flicker amplitudes between mice that received the medium 8E8 vg dose and WT or heterozygous control mice. At 1 MPI (2 MO), treatment with the 8E8 vg dose restored ERGs to near-WT amplitudes. (F) Waveforms from an eye that received the medium 8E8 dose and a WT eye. MO, months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: rAAV2tYF subretinal gene therapy improves and sustains cone photoreceptor function in Rs1 -KO mice treated with the medium dose of 8E8 vg/eye over the course of 12 months. (A) Cone-dependent retinal function, assessed through light-adapted (LA) electroretinography after subjecting the eyes to a 3.0 cd·s/m 2 bright flash. Eyes treated with the 8E8 vg dose consistently displayed significantly greater b-wave amplitudes compared to sham-treated eyes and untreated eyes over 1 year. Eyes treated with 8E9 showed significance compared to the sham-treated eyes only at the experimental endpoint. (B) Representative LA waveforms were shown for eyes treated with different doses, as well as for sham-treated, and untreated eyes at 3 MPI and 7 MPI, respectively. (C) Another metric used to evaluate cone-dependent retinal function was the 5 Hz flicker test, which consistently showed that 8E8 vg dose-treated eyes outperformed sham-treated and untreated eyes throughout the treatment duration. Neither 8E9 nor 8E7 vg doses showed significance compared to the sham-treated eyes over the treatment course. (D) When subjected to a 5 Hz flicker, cones in treated eyes elicited higher amplitudes at 3 MPI, which was not observed in the sham-treated eyes or the untreated contralateral eyes. Over the course of 7 months, treated eyes retained significantly better amplitudes in cone ERGs. (E) Comparison of ERG flicker amplitudes between mice that received the medium 8E8 vg dose and WT or heterozygous control mice. At 1 MPI (2 MO), treatment with the 8E8 vg dose restored ERGs to near-WT amplitudes. (F) Waveforms from an eye that received the medium 8E8 dose and a WT eye. MO, months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Techniques: Comparison, Control

Subretinal gene therapy of Rs1 -KO delays loss of retinal function in standard combined response ERG. (A) Combined rod-cone function was measured by 3.0 cd·s/m 2 bright flash under dark-adapted conditions, at 1, 2, 3, 5, 7 and 12 MPI. The 8E8 vg dose treated eyes had higher b-wave amplitudes compared to diluent-injected eyes and untreated contralateral eyes. (B,C) Representative waveforms of the 3.0 cd·s/m 2 bright flash SCR ERG test are shown for all vector doses, diluent-injected eyes, and untreated eyes at 3 MPI and 7 MPI, respectively. The 8E8 dose had higher amplitudes at 3 MPI and at 7MPI. Waveforms from the same mice were used at each time point. (D) An elevated b/a ratio was observed in the 8E8 vg dose-treated eyes until 2 MPI time point and in 8E9 vg dose treated eyes until 5 MPI. (E) SCR a-wave amplitudes of eyes treated with different doses of rAAV2tYF gene therapy vector, sham-treated and untreated control mice. (F) Rod and cone function comparison between the medium 8E8 vg dose and WT or heterozygous control mice at 2 and 6 MO. MO, months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Subretinal gene therapy of Rs1 -KO delays loss of retinal function in standard combined response ERG. (A) Combined rod-cone function was measured by 3.0 cd·s/m 2 bright flash under dark-adapted conditions, at 1, 2, 3, 5, 7 and 12 MPI. The 8E8 vg dose treated eyes had higher b-wave amplitudes compared to diluent-injected eyes and untreated contralateral eyes. (B,C) Representative waveforms of the 3.0 cd·s/m 2 bright flash SCR ERG test are shown for all vector doses, diluent-injected eyes, and untreated eyes at 3 MPI and 7 MPI, respectively. The 8E8 dose had higher amplitudes at 3 MPI and at 7MPI. Waveforms from the same mice were used at each time point. (D) An elevated b/a ratio was observed in the 8E8 vg dose-treated eyes until 2 MPI time point and in 8E9 vg dose treated eyes until 5 MPI. (E) SCR a-wave amplitudes of eyes treated with different doses of rAAV2tYF gene therapy vector, sham-treated and untreated control mice. (F) Rod and cone function comparison between the medium 8E8 vg dose and WT or heterozygous control mice at 2 and 6 MO. MO, months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Techniques: Injection, Plasmid Preparation, Control, Comparison

Subretinal gene therapy of Rs1 -KO with a dose of 8E8 vg/eye delays loss of rod photoreceptor function. (A) Rod function was measured by 0.01 cd·s/m 2 dim flash after DA. Notably, at 1, 2, and 3 MPI, some vector-treated eyes demonstrated a normalization of the hyper-normal a-wave seen in the Rs1 -KO mouse model (as shown in ). (B) B-wave amplitudes of the 0.01 cd·s/m 2 dim flash after DA showing the significantly increased b-wave amplitudes of the 8E8 vg dose compared to the sham-treated and untreated eyes. (C,D) Waveform comparisons of the 0.01 cd·s/m 2 dim flash test were made between all vector doses, diluent-injected eyes, and untreated eyes at 3 MPI and 7 MPI, respectively. (E) Rod function comparison between the medium 8E8 vg dose and WT or heterozygous control mice (WT) at 2 and 6 MO. MO: months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p ≤ 0.005, *** p ≤ 0.0005, and **** p < 0.0001.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Subretinal gene therapy of Rs1 -KO with a dose of 8E8 vg/eye delays loss of rod photoreceptor function. (A) Rod function was measured by 0.01 cd·s/m 2 dim flash after DA. Notably, at 1, 2, and 3 MPI, some vector-treated eyes demonstrated a normalization of the hyper-normal a-wave seen in the Rs1 -KO mouse model (as shown in ). (B) B-wave amplitudes of the 0.01 cd·s/m 2 dim flash after DA showing the significantly increased b-wave amplitudes of the 8E8 vg dose compared to the sham-treated and untreated eyes. (C,D) Waveform comparisons of the 0.01 cd·s/m 2 dim flash test were made between all vector doses, diluent-injected eyes, and untreated eyes at 3 MPI and 7 MPI, respectively. (E) Rod function comparison between the medium 8E8 vg dose and WT or heterozygous control mice (WT) at 2 and 6 MO. MO: months old, WT: wild-type or heterozygous mice, µV: Microvolt, * p < 0.05, ** p ≤ 0.005, *** p ≤ 0.0005, and **** p < 0.0001.

Techniques: Plasmid Preparation, Injection, Comparison, Control

Eyes treated with the 8E8 vg dose exhibit superior cone rescue at 14 months old compared to sham-treated and untreated eyes. (A–E) Visualization of cone outer segments using peanut agglutinin. Retinal sections collected from all vector treated eyes, diluent-treated eyes, and untreated Rs1 -KO eyes at 14 months of age were processed and stained with DAPI (blue) and PNA (red) to visualize cone outer segments. (F) Quantification of cone outer segments per 100 μm of the retina indicates that there is a significant preservation of cones in eyes treated with the 8E8 vg dose compared to eyes that received sham treatment and untreated Rs1 -KO eyes. Eyes treated with all doses of the vector, as well as diluent-treated eyes, had significantly more cones than untreated eyes. Quantification was performed by three individuals masked to treatment groups. OS, outer segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer. * p < 0.05, ** p < 0.005. Scale bar, 50 μm.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Eyes treated with the 8E8 vg dose exhibit superior cone rescue at 14 months old compared to sham-treated and untreated eyes. (A–E) Visualization of cone outer segments using peanut agglutinin. Retinal sections collected from all vector treated eyes, diluent-treated eyes, and untreated Rs1 -KO eyes at 14 months of age were processed and stained with DAPI (blue) and PNA (red) to visualize cone outer segments. (F) Quantification of cone outer segments per 100 μm of the retina indicates that there is a significant preservation of cones in eyes treated with the 8E8 vg dose compared to eyes that received sham treatment and untreated Rs1 -KO eyes. Eyes treated with all doses of the vector, as well as diluent-treated eyes, had significantly more cones than untreated eyes. Quantification was performed by three individuals masked to treatment groups. OS, outer segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer. * p < 0.05, ** p < 0.005. Scale bar, 50 μm.

Techniques: Plasmid Preparation, Staining, Preserving

Medium dose (8E8 vg) of subretinal rAAV2tYF gene therapy shows improvement in functional vision. (A) Under normal fluorescent ceiling light, Rs1- KO mice treated with 8E8 vg dosage of the rAAV2tYF subretinal gene therapy took significantly less time to locate the platform compared to untreated Rs1- KO mice but not compared to the sham-treated mice at both time points (4–6 and 9–11 MO). (B) In dark testing conditions (dim red lighting), mice treated with the 8E8 vg dose of the gene therapy took significantly less time to locate the platform compared to untreated Rs1- KO mice at younger ages but not at older ages. No statistical difference was observed for any of the vector treated eyes compared to the sham-treated eyes. (C) Confidence intervals were employed to show the data spread. No statistically significant difference or clear trend towards improvement at older age compared to the untreated eyes was observed. MO: months old, * p < 0.05.

Journal: Frontiers in Medicine

Article Title: The dose-response relationship of subretinal gene therapy with rAAV2tYF-CB-h RS1 in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1304819

Figure Lengend Snippet: Medium dose (8E8 vg) of subretinal rAAV2tYF gene therapy shows improvement in functional vision. (A) Under normal fluorescent ceiling light, Rs1- KO mice treated with 8E8 vg dosage of the rAAV2tYF subretinal gene therapy took significantly less time to locate the platform compared to untreated Rs1- KO mice but not compared to the sham-treated mice at both time points (4–6 and 9–11 MO). (B) In dark testing conditions (dim red lighting), mice treated with the 8E8 vg dose of the gene therapy took significantly less time to locate the platform compared to untreated Rs1- KO mice at younger ages but not at older ages. No statistical difference was observed for any of the vector treated eyes compared to the sham-treated eyes. (C) Confidence intervals were employed to show the data spread. No statistically significant difference or clear trend towards improvement at older age compared to the untreated eyes was observed. MO: months old, * p < 0.05.

Techniques: Functional Assay, Plasmid Preparation

Relationship between RS1 expression level and clinicopathological variables in LUAD patients

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Relationship between RS1 expression level and clinicopathological variables in LUAD patients

Article Snippet: In the Western blot, the primary antibody was RS1 polyclonal antibody (1:1000, H00006247‐B01P; Novus), and the secondary antibody was IgG‐HRP (1:3000, bs‐0295G‐HRP; Bioss).

Techniques: Expressing, Mutagenesis

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Primer sequences for RT‐PCR

Techniques:

Expression levels of RS1 gene in lung adenocarcinoma (LUAD). (a) Expression difference of RS1 gene between tumor tissues and adjacent tissues of LUAD samples in the TCGA database. The longitudinal axis represents the relative expression level of mRNA. (b) Expression difference of RS1 gene between 56 LUAD tissues and paired adjacent tissues in the TCGA database. (c) RS1 expression levels at different TNM stages. (d) RS1 expression levels at different T stages. (e) RS1 expression levels at different N stages. (f) Expression difference of RS1 gene between tumor and adjacent tissues of LUAD samples in GEPIA database. (g) Different expression levels between tumor and paired adjacent tissues in the GSE32863 dataset. (h) Different expression levels between tumor tissues and paired adjacent tissues in GSE115002 dataset. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Expression levels of RS1 gene in lung adenocarcinoma (LUAD). (a) Expression difference of RS1 gene between tumor tissues and adjacent tissues of LUAD samples in the TCGA database. The longitudinal axis represents the relative expression level of mRNA. (b) Expression difference of RS1 gene between 56 LUAD tissues and paired adjacent tissues in the TCGA database. (c) RS1 expression levels at different TNM stages. (d) RS1 expression levels at different T stages. (e) RS1 expression levels at different N stages. (f) Expression difference of RS1 gene between tumor and adjacent tissues of LUAD samples in GEPIA database. (g) Different expression levels between tumor and paired adjacent tissues in the GSE32863 dataset. (h) Different expression levels between tumor tissues and paired adjacent tissues in GSE115002 dataset. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Techniques: Expressing

RS1 gene is an independent prognostic indicator for lung adenocarcinoma (LUAD). (a) Kaplan–Meier survival analysis for TCGA dataset showed poor overall survival (OS) in samples with low expression of RS1. (b) Kaplan–Meier survival analysis for GSE37745 dataset. (c) A forest map of six factors included in multivariate Cox regression analysis. Compared with reference, HR >1 represents a higher risk of death, and HR <1 represents a lower risk of death.

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: RS1 gene is an independent prognostic indicator for lung adenocarcinoma (LUAD). (a) Kaplan–Meier survival analysis for TCGA dataset showed poor overall survival (OS) in samples with low expression of RS1. (b) Kaplan–Meier survival analysis for GSE37745 dataset. (c) A forest map of six factors included in multivariate Cox regression analysis. Compared with reference, HR >1 represents a higher risk of death, and HR <1 represents a lower risk of death.

Techniques: Expressing

Immune infiltration of lung adenocarcinoma (LUAD) patients in the high and low RS1 expression groups. (a) Infiltration rate of 22 immune cells in all patients. (b) A correlation matrix of 22 immune infiltration proportions. Red represents positive correlation and blue represents negative correlation, with darker colors meaning stronger correlation. (c) The infiltration difference of immune cells in the RS1 high and low expression groups. The vertical axis represents the relative infiltration proportions of immune cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Immune infiltration of lung adenocarcinoma (LUAD) patients in the high and low RS1 expression groups. (a) Infiltration rate of 22 immune cells in all patients. (b) A correlation matrix of 22 immune infiltration proportions. Red represents positive correlation and blue represents negative correlation, with darker colors meaning stronger correlation. (c) The infiltration difference of immune cells in the RS1 high and low expression groups. The vertical axis represents the relative infiltration proportions of immune cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Techniques: Expressing

GSEA enriched KEGG pathways in high and low expression of RS1

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: GSEA enriched KEGG pathways in high and low expression of RS1

Techniques: Expressing

Gene set enrichment analysis (GSEA) showed the top six pathways enriched in the high RS1 expression group. (a) Cardiac muscle contraction pathway, (b) vascular smooth muscle contraction pathway, (c) renin angiotensin system pathway, (d) hypertrophic cardiomyopathy (HCM), (e) GnRH signaling pathway and (f) differentiated cardiomyopathy pathway

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) showed the top six pathways enriched in the high RS1 expression group. (a) Cardiac muscle contraction pathway, (b) vascular smooth muscle contraction pathway, (c) renin angiotensin system pathway, (d) hypertrophic cardiomyopathy (HCM), (e) GnRH signaling pathway and (f) differentiated cardiomyopathy pathway

Techniques: Expressing

RS1 expression in cell lines. (a) RS1 mRNA was downregulated in lung adenocarcinoma (LUAD) cells. *** p < 0.001 versus NCI‐H441; ### p < 0.001 versus NCI‐H1975. (b) Expression of RS1 protein in LUAD cells. (c) Comparison of gray values in WB. *** p < 0.001 versus NCI‐H441; ### p < 0.001 versus NCI‐H1975.

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: RS1 expression in cell lines. (a) RS1 mRNA was downregulated in lung adenocarcinoma (LUAD) cells. *** p < 0.001 versus NCI‐H441; ### p < 0.001 versus NCI‐H1975. (b) Expression of RS1 protein in LUAD cells. (c) Comparison of gray values in WB. *** p < 0.001 versus NCI‐H441; ### p < 0.001 versus NCI‐H1975.

Techniques: Expressing, Comparison

Representative images of RS1 protein expressed in clinical samples

Journal: Thoracic Cancer

Article Title: RS1 gene is a novel prognostic biomarker for lung adenocarcinoma

doi: 10.1111/1759-7714.14471

Figure Lengend Snippet: Representative images of RS1 protein expressed in clinical samples

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