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Chem Impex International 88930 14 7 chem impex 04461
88930 14 7 Chem Impex 04461, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/04461/N-Methyl-D-valine+hydrochloride/10__1038_slash_s41557___018___0033___8____41557_2018_33_MOESM1_ESM-225-89-90
Average 96 stars, based on 1 article reviews

88930 14 7 chem impex 04461 - by Bioz Stars, 2026-09

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Article Title: Second-Generation DNA-Templated Macrocycle Libraries for the Discovery of Bioactive Small Molecules
Article Snippet: Chemical building blocks 2 used for the secondgeneration library parent amino acid CAS number commercial source 2A Disoglutamine (HCl) 19522408 ChemImpex 05966 2B OmethylLserine 32620114 Astatech F10843 2C 4(methylamino)butanoic acid 1119488 Astatech AB7427 2D D2carbamoylphenylalanine 1217613529 ChemImpex 16773 2E 1(aminomethyl)cyclopropanecarboxylic acid 139126457 AldrichCPR CDS015451100MG 2F NmethylDphenylalanine 56564524 Alfa Aesar H65675 2G Laspartic acid αmethyl ester 17812327 ChemImpex 02695 2H (2methylaminoethoxy)acetic acid 98137587 Oakwood 0952191g 2I D3cyclobutylalanine 174266003 Astatech 59421 2J 3cyclohexylLalanine 27527055 ChemImpex 02560 2K Dproline 344252 SigmaAldrich 858919 2L NmethylLthreonine (HCl) 2812284 ChemImpex 09616 2M Nmethyl DValine (HCl) 88930147 ChemImpex 04461 2N (1R,3S)3aminocyclopentane carboxylic acid 71830085 ChemImpex 15488 2O L2furylalanine 121786310 ChemImpex 07442 2P transL4hydroxyproline 51354 ChemImpex 00185 2Q 1aminocyclobutanecarboxylic acid 22264502 Oakwood 0664721g 2R 5amino4oxopentanoic acid 106605 Astatech 27973 2S 1aminocyclopropane1carboxylic acid 22059218 ChemImpex 07075 2T Nmethylglycine 107971 ChemImpex 01319 Supplementary Table 22.

Article Title: Second-generation DNA-templated macrocycle libraries for the discovery of bioactive small molecules
Article Snippet: Chemical building blocks 2 used for the second-generation library parent amino acid CAS number commercial source 2A D-isoglutamine (HCl) 19522-40-8 Chem-Impex 05966 2B O-methyl-L-serine 32620-11-4 Astatech F10843 2C 4-(methylamino)butanoic acid 1119-48-8 Astatech AB7427 2D D-2-carbamoylphenylalanine 1217613-52-9 Chem-Impex 16773 2E 1-(aminomethyl)cyclopropanecarboxylic acid 139126-45-7 AldrichCPR CDS015451-100MG 2F N-methyl-D-phenylalanine 56564-52-4 Alfa Aesar H65675 2G L-aspartic acid α-methyl ester 17812-32-7 Chem-Impex 02695 2H (2-methylamino-ethoxy)-acetic acid 98137-58-7 Oakwood 095219-1g 2I D-3-cyclobutylalanine 174266-00-3 Astatech 59421 2J 3-cyclohexyl-L-alanine 27527-05-5 Chem-Impex 02560 2K D-proline 344-25-2 Sigma-Aldrich 858919 2L N-methyl-L-threonine (HCl) 2812-28-4 Chem-Impex 09616 2M N-methyl- D-Valine (HCl) 88930-14-7 Chem-Impex 04461 2N (1R,3S)-3-aminocyclopentane carboxylic acid 71830-08-5 Chem-Impex 15488 2O L-2-furylalanine 121786-31-0 Chem-Impex 07442 2P trans-L-4-hydroxyproline 51-35-4 Chem-Impex 00185 2Q 1-aminocyclobutanecarboxylic acid 22264-50-2 Oakwood 066472-1g 2R 5-amino-4-oxopentanoic acid 106-60-5 Astatech 27973 2S 1-aminocyclopropane-1-carboxylic acid 22059-21-8 Chem-Impex 07075 2T N-methylglycine 107-97-1 Chem-Impex 01319 Supplementary Table 22.

Image Search Results

(A) Immunostaining of PROM1, ARL13B, and VSX2 in optic-cup-like structures at D24 of C1-ROs differentiation. Antibody specificity was confirmed using negative controls. Magnified regions are indicated by white dashed boxes. Scale bars: 100 µm (overview) and 20 µm (magnified view). (B) Immunostaining of RCVRN and PROM1 in C1-ROs at D60 and D90. Scale bar, 40 μm. (C) Expression and distribution of rod (RHOD + ) and cone (ARR3 + ) in C1-ROs at D120, D150, and D210. Scale bar, 40 μm.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Immunostaining of PROM1, ARL13B, and VSX2 in optic-cup-like structures at D24 of C1-ROs differentiation. Antibody specificity was confirmed using negative controls. Magnified regions are indicated by white dashed boxes. Scale bars: 100 µm (overview) and 20 µm (magnified view). (B) Immunostaining of RCVRN and PROM1 in C1-ROs at D60 and D90. Scale bar, 40 μm. (C) Expression and distribution of rod (RHOD + ) and cone (ARR3 + ) in C1-ROs at D120, D150, and D210. Scale bar, 40 μm.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Immunostaining, Expressing

(A and B) Immunostaining of PROM1 during C1-ROs differentiation, co-labeled with ARL13B (cilia marker) or PRPH2 (OS marker). Nuclei are counterstained with DAPI. Time points: D40, D60, D90, D120, and D150. Scale bar, 20 µm. (C) Schematic diagram of whole-ROs imaging. The top view illustrates the coverslip (black square), the hydrophobic barrier drawn with a PAP pen (pink square), the antifade mounting medium (blue area), and the RO (yellow sphere). The frontal view shows the red laser and the confocal microscope objective positioned beneath the coverslip. (D) Whole-mount staining of D300 C1-ROs showing PROM1 expression in PRPH2 + OS-like structures and ARR3 + cones. Scale bar, 10 µm. (E) Co-localization of PROM1 with Rhodopsin (rods), L/M-opsin (L/M-cones), and S-opsin (S-cones) in D300 C1-ROs. Scale bar, 2 μm. (F and G) Confocal immunostaining of PROM1, MCT1, and ZO-1 in C1 hiPSC-RPE cells cultured on Transwells. Y–Z projections illustrate subcellular localization. (H and I) Schematics showing alternative splicing patterns of PROM1 in exon 4 (H) and exons 25–26 (I) in C1-hiPSCs versus C1-ROs. (J) Western blot analysis of PROM1 expression and N-glycosylation in hiPSCs, hiPSC-RPE cells, and ROs. C1 and C2 represent different normal individuals. “F” indicates Glycopeptidase F-treated samples; “−” denotes untreated controls. (K and L) Diagrams depicting glycosylated PROM1 in the plasma membrane of hiPSCs (K) and photoreceptors (L).

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A and B) Immunostaining of PROM1 during C1-ROs differentiation, co-labeled with ARL13B (cilia marker) or PRPH2 (OS marker). Nuclei are counterstained with DAPI. Time points: D40, D60, D90, D120, and D150. Scale bar, 20 µm. (C) Schematic diagram of whole-ROs imaging. The top view illustrates the coverslip (black square), the hydrophobic barrier drawn with a PAP pen (pink square), the antifade mounting medium (blue area), and the RO (yellow sphere). The frontal view shows the red laser and the confocal microscope objective positioned beneath the coverslip. (D) Whole-mount staining of D300 C1-ROs showing PROM1 expression in PRPH2 + OS-like structures and ARR3 + cones. Scale bar, 10 µm. (E) Co-localization of PROM1 with Rhodopsin (rods), L/M-opsin (L/M-cones), and S-opsin (S-cones) in D300 C1-ROs. Scale bar, 2 μm. (F and G) Confocal immunostaining of PROM1, MCT1, and ZO-1 in C1 hiPSC-RPE cells cultured on Transwells. Y–Z projections illustrate subcellular localization. (H and I) Schematics showing alternative splicing patterns of PROM1 in exon 4 (H) and exons 25–26 (I) in C1-hiPSCs versus C1-ROs. (J) Western blot analysis of PROM1 expression and N-glycosylation in hiPSCs, hiPSC-RPE cells, and ROs. C1 and C2 represent different normal individuals. “F” indicates Glycopeptidase F-treated samples; “−” denotes untreated controls. (K and L) Diagrams depicting glycosylated PROM1 in the plasma membrane of hiPSCs (K) and photoreceptors (L).

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Immunostaining, Labeling, Marker, Imaging, Microscopy, Staining, Expressing, Cell Culture, Alternative Splicing, Western Blot, Glycoproteomics, Clinical Proteomics, Membrane

Immunostaining shows the expression and distribution of PROM1 in both ROs at multiple time points. PRPH2 was used to label OS-like structures. C1-ROs and C2-ROs were derived from two independent wild-type hiPSC lines. Scale bar, 20 μm.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: Immunostaining shows the expression and distribution of PROM1 in both ROs at multiple time points. PRPH2 was used to label OS-like structures. C1-ROs and C2-ROs were derived from two independent wild-type hiPSC lines. Scale bar, 20 μm.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Immunostaining, Expressing, Derivative Assay

(A) Bright-field image of C1 hiPSC-RPE cells cultured on Transwells for 6 weeks. Scale bar, 100 μm. (B) Immunostaining of PROM1 together with MCT1 and ZO-1 in C1 hiPSC-RPE cells, corresponding to and . Scale bar, 40 μm. The negative control confirms specificity. Scale bar, 40 μm.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Bright-field image of C1 hiPSC-RPE cells cultured on Transwells for 6 weeks. Scale bar, 100 μm. (B) Immunostaining of PROM1 together with MCT1 and ZO-1 in C1 hiPSC-RPE cells, corresponding to and . Scale bar, 40 μm. The negative control confirms specificity. Scale bar, 40 μm.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Cell Culture, Immunostaining, Negative Control

(A) Schematic of the PROM1 genomic structure and ten alternatively spliced mRNA isoforms; transcript variant 1 (NM_006017.3) is the longest, containing all 28 exons. Major splicing differences at exon 4 and exons 25–26 are highlighted. (B) Diagram of PROM1 mRNA showing primer binding sites used for RT-PCR. (C) Bidirectional Sanger sequencing of PROM1 splice variants involving exon 4 and exons 25–26 in C1-hiPSCs and C1-ROs; sequencing directions are indicated by black arrows. (D and E) TA cloning and Sanger sequencing of PROM1 transcripts spanning exons 3–12 (D) and exons 24–27 (E) from C1-ROs.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Schematic of the PROM1 genomic structure and ten alternatively spliced mRNA isoforms; transcript variant 1 (NM_006017.3) is the longest, containing all 28 exons. Major splicing differences at exon 4 and exons 25–26 are highlighted. (B) Diagram of PROM1 mRNA showing primer binding sites used for RT-PCR. (C) Bidirectional Sanger sequencing of PROM1 splice variants involving exon 4 and exons 25–26 in C1-hiPSCs and C1-ROs; sequencing directions are indicated by black arrows. (D and E) TA cloning and Sanger sequencing of PROM1 transcripts spanning exons 3–12 (D) and exons 24–27 (E) from C1-ROs.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Variant Assay, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Sequencing, TA Cloning

(A) Bright-field images of ROs at D270. Scale bar, 100 µm. (B) Immunostaining shows expression of PROM1, ARL13B, and PRPH2 across differentiation timepoints (D40–D270) in Het-ROs and Hom-ROs. Scale bar, 20 µm. (C) PROM1 expression in ROs was assessed using PROM1-N and PROM1-C antibodies. (D) Original Western blot images corresponding to .

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Bright-field images of ROs at D270. Scale bar, 100 µm. (B) Immunostaining shows expression of PROM1, ARL13B, and PRPH2 across differentiation timepoints (D40–D270) in Het-ROs and Hom-ROs. Scale bar, 20 µm. (C) PROM1 expression in ROs was assessed using PROM1-N and PROM1-C antibodies. (D) Original Western blot images corresponding to .

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Immunostaining, Expressing, Western Blot

(A) Pedigree of the PROM1-IRD family. The proband (F2-2) indicated by a black arrow. (B-D) Multimodal retinal imaging of the 35-year-old proband, including fundus photographs(B), FFA (C) and SD-OCT (D). RE, right eye; LE, left eye. (E) Bright-field images of UCs and hiPSCs from the proband (Hom) and his mother (Het). Scale bar, 100 μm. (F and H) Western blot showing PROM1 protein levels in UCs and hiPSCs. β-actin serves as the loading control. (G) qRT-PCR analysis of PROM1 mRNA expression in Het-hiPSCs (n=3) and Hom-hiPSCs (n=3). Error bars represent mean ± S.E.M.; ***P < 0.001. (I and J) TA cloning and sequencing of PROM1 cDNA from Hom-hiPSCs. Among 7 clones, 5 exhibited an 82 bp deletion (Mut type I), and 2 harbored the c.619 G>T point mutation (Mut type II). The corresponding sequence is detailed in (J).

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Pedigree of the PROM1-IRD family. The proband (F2-2) indicated by a black arrow. (B-D) Multimodal retinal imaging of the 35-year-old proband, including fundus photographs(B), FFA (C) and SD-OCT (D). RE, right eye; LE, left eye. (E) Bright-field images of UCs and hiPSCs from the proband (Hom) and his mother (Het). Scale bar, 100 μm. (F and H) Western blot showing PROM1 protein levels in UCs and hiPSCs. β-actin serves as the loading control. (G) qRT-PCR analysis of PROM1 mRNA expression in Het-hiPSCs (n=3) and Hom-hiPSCs (n=3). Error bars represent mean ± S.E.M.; ***P < 0.001. (I and J) TA cloning and sequencing of PROM1 cDNA from Hom-hiPSCs. Among 7 clones, 5 exhibited an 82 bp deletion (Mut type I), and 2 harbored the c.619 G>T point mutation (Mut type II). The corresponding sequence is detailed in (J).

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Imaging, Western Blot, Control, Quantitative RT-PCR, Expressing, TA Cloning, Sequencing, Clone Assay, Mutagenesis

(A) Detection of the PROM1 mutation in members of the PROM1-IRD pedigree. Red arrow indicates the mutation site. (B and C) Sanger sequencing analysis of the PROM1 mutation in cDNA from UCs and hiPSCs. The left panel shows the beginning of overlapping peaks at position c.548 (yellow arrow), while the right panel indicates the disappearance of these overlaps at position c.630 (red arrow).

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Detection of the PROM1 mutation in members of the PROM1-IRD pedigree. Red arrow indicates the mutation site. (B and C) Sanger sequencing analysis of the PROM1 mutation in cDNA from UCs and hiPSCs. The left panel shows the beginning of overlapping peaks at position c.548 (yellow arrow), while the right panel indicates the disappearance of these overlaps at position c.630 (red arrow).

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Mutagenesis, Sequencing

(A and B) Original Western blot images correspond to and . (C) PROM1 expression in UCs and hiPSCs was assessed using the PROM1-N antibody, which specifically recognizes the N-terminal region (aa 20– 108) of PROM1. α-Tubulin serves as loading control.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A and B) Original Western blot images correspond to and . (C) PROM1 expression in UCs and hiPSCs was assessed using the PROM1-N antibody, which specifically recognizes the N-terminal region (aa 20– 108) of PROM1. α-Tubulin serves as loading control.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Western Blot, Expressing, Control

(A) Immunostaining of PROM1 and PRPH2 in D150 ROs. Regions within white dashed boxes are shown enlarged. Scale bars: 40 μm (overview) and 10 μm (magnified). (B and C) Quantitative analysis of OS-like structures lengths in C1-ROs (n=444), Het-ROs (n=873), and Hom-ROs (n=396). The lengths were categorized into four groups: 0–1 μm, 1–2 μm, 2–3 μm, and >3 μm. (D) Quantification of OS-like structures per 40×40 μm². C1-ROs (n=4), Het-ROs (n=5), Hom-ROs (n=4). (E) TEM images of OS-like structures in D180 C1-ROs and Hom-ROs. Scale bars: 20 μm (left), 5 μm (right). (F) Top 10 significantly enriched KEGG pathways comparing D150 C1-ROs and Hom-ROs. (G) Heatmap of DEGs in the PI3K-Akt signaling pathway across all groups. (H and I) Western blot showing PROM1 protein levels in D150 ROs (n=4). β-actin as control. (J) RNA-seq analysis showing FPKM values of PROM1 in D150 ROs (n=3). (K) ONL thickness quantification in D150 ROs based on DAPI staining. C1-ROs (n=8), Het-ROs (n=6), Hom-ROs (n=3). (L and M) TUNEL staining for photoreceptors apoptosis in D150 ROs. Apoptotic cells per 0.01 mm² quantified. C1-ROs (n=3), Het-ROs (n=3), and Hom-ROs (n=6); Scale bars, 40 μm. Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001 .

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Immunostaining of PROM1 and PRPH2 in D150 ROs. Regions within white dashed boxes are shown enlarged. Scale bars: 40 μm (overview) and 10 μm (magnified). (B and C) Quantitative analysis of OS-like structures lengths in C1-ROs (n=444), Het-ROs (n=873), and Hom-ROs (n=396). The lengths were categorized into four groups: 0–1 μm, 1–2 μm, 2–3 μm, and >3 μm. (D) Quantification of OS-like structures per 40×40 μm². C1-ROs (n=4), Het-ROs (n=5), Hom-ROs (n=4). (E) TEM images of OS-like structures in D180 C1-ROs and Hom-ROs. Scale bars: 20 μm (left), 5 μm (right). (F) Top 10 significantly enriched KEGG pathways comparing D150 C1-ROs and Hom-ROs. (G) Heatmap of DEGs in the PI3K-Akt signaling pathway across all groups. (H and I) Western blot showing PROM1 protein levels in D150 ROs (n=4). β-actin as control. (J) RNA-seq analysis showing FPKM values of PROM1 in D150 ROs (n=3). (K) ONL thickness quantification in D150 ROs based on DAPI staining. C1-ROs (n=8), Het-ROs (n=6), Hom-ROs (n=3). (L and M) TUNEL staining for photoreceptors apoptosis in D150 ROs. Apoptotic cells per 0.01 mm² quantified. C1-ROs (n=3), Het-ROs (n=3), and Hom-ROs (n=6); Scale bars, 40 μm. Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001 .

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Immunostaining, Western Blot, Control, RNA Sequencing, Staining, TUNEL Assay

(A) D90 C1-ROs was infected with AAV2-CMV-GFP , AAV7m8-CMV-GFP or AAV8-CMV-GFP (1×10 11 vg/RO). Bright-field and fluorescence images taken before and after infection. Scale bar, 500 µm. (B) Schematic of pAAV-CRXp-hPROM1 construct. (C) Experimental outline of the AAV -mediated gene augmentation in Hom-ROs. (D) Immunostaining for PROM1 and PRPH2 in D120 ROs. Hom-ROs infected at D90 is denoted Hom-ROs/AAV7m8. Regions within dashed boxes were magnified on the right. Scale bars: 40 µm (left) and 10 µm (right). (E) Proportions of PROM1 + /PRPH2 + versus PROM1 - /PRPH2 + OS-like structures in D90+30 Hom-ROs/AAV7m8. (F and G) Quantitative analysis of OS-like structures lengths in C1-ROs (n=486), Het-ROs (n=358), Hom-ROs (n=433), Hom-ROs/AAV7m8 (PROM1 + /PRPH2 + ; n=297), and Hom-ROs/AAV7m8 (PROM1 - /PRPH2 + ; n=242). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); ***P < 0.001 .

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) D90 C1-ROs was infected with AAV2-CMV-GFP , AAV7m8-CMV-GFP or AAV8-CMV-GFP (1×10 11 vg/RO). Bright-field and fluorescence images taken before and after infection. Scale bar, 500 µm. (B) Schematic of pAAV-CRXp-hPROM1 construct. (C) Experimental outline of the AAV -mediated gene augmentation in Hom-ROs. (D) Immunostaining for PROM1 and PRPH2 in D120 ROs. Hom-ROs infected at D90 is denoted Hom-ROs/AAV7m8. Regions within dashed boxes were magnified on the right. Scale bars: 40 µm (left) and 10 µm (right). (E) Proportions of PROM1 + /PRPH2 + versus PROM1 - /PRPH2 + OS-like structures in D90+30 Hom-ROs/AAV7m8. (F and G) Quantitative analysis of OS-like structures lengths in C1-ROs (n=486), Het-ROs (n=358), Hom-ROs (n=433), Hom-ROs/AAV7m8 (PROM1 + /PRPH2 + ; n=297), and Hom-ROs/AAV7m8 (PROM1 - /PRPH2 + ; n=242). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); ***P < 0.001 .

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Infection, Fluorescence, Construct, Immunostaining

Hom-ROs infected with AAV7m8-CRXp-hPROM1 was analyzed for PROM1 expression at D90+30 (A), D90+60 (B) and D150+80 (C). Magnified regions indicated by white dashed boxes.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: Hom-ROs infected with AAV7m8-CRXp-hPROM1 was analyzed for PROM1 expression at D90+30 (A), D90+60 (B) and D150+80 (C). Magnified regions indicated by white dashed boxes.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Infection, Expressing

(A) Schematic of Prom1 gene knockout strategy in mice. (B) SD-OCT images of 3-month-old mice retinas. Layers indicated: GCL, INL, ONL, IS and OS. Scale bar, 100 µm. (C–E) H&E staining of Prom1 +/+ , Prom1 +/- , and Prom1 -/- retinas. Quantification of ONL and IS/OS thickness in (D) and (E), respectively. n=3 per group. (F) Schematic of subretinal AAV injection. (G) Bright-field and GFP images of retina one-week post-injection of AAV8-CMV-GFP in Prom1 -/- mice. Scale bar, 200 µm. (H) Western blot analysis of exogenous human PROM1 and endogenous mouse PROM1 expression in retinas. Three biological replicates labeled 1#, 2#, and 3#. (I) Immunostaining showing distribution of hPROM1 and GFP-labeled photoreceptors in Prom1 -/- retina post co-injection of AAV8-CRXp-hPROM1 and AAV8-CMV-GFP (1:10). Scale bar, 40 µm. (J) Immunostaining of PROM1 in the prom1 -/- mice with or without AAV8-CRXp-hPROM1 infection. Scale bar, 50 µm. (K and L) H&E staining and quantification of ONL and IS/OS thickness 2 months after AAV injection (n=3). Scale bar, 40 µm. (M–O) fERG recordings showing representative Scoptoic 3.0 waveforms (M) and quantification of a- and b-wave amplitudes (N and O). A stratified statistical analysis was performed to specifically assess gene therapy efficacy. Scale bar, 100 µV, 25 ms. Groups: Prom1 +/+ (n=3), Prom1 +/- (n=3), Prom1 -/- (n=7), and Prom1 -/- +AAV8-hPROM1 (n=8). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001 .

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) Schematic of Prom1 gene knockout strategy in mice. (B) SD-OCT images of 3-month-old mice retinas. Layers indicated: GCL, INL, ONL, IS and OS. Scale bar, 100 µm. (C–E) H&E staining of Prom1 +/+ , Prom1 +/- , and Prom1 -/- retinas. Quantification of ONL and IS/OS thickness in (D) and (E), respectively. n=3 per group. (F) Schematic of subretinal AAV injection. (G) Bright-field and GFP images of retina one-week post-injection of AAV8-CMV-GFP in Prom1 -/- mice. Scale bar, 200 µm. (H) Western blot analysis of exogenous human PROM1 and endogenous mouse PROM1 expression in retinas. Three biological replicates labeled 1#, 2#, and 3#. (I) Immunostaining showing distribution of hPROM1 and GFP-labeled photoreceptors in Prom1 -/- retina post co-injection of AAV8-CRXp-hPROM1 and AAV8-CMV-GFP (1:10). Scale bar, 40 µm. (J) Immunostaining of PROM1 in the prom1 -/- mice with or without AAV8-CRXp-hPROM1 infection. Scale bar, 50 µm. (K and L) H&E staining and quantification of ONL and IS/OS thickness 2 months after AAV injection (n=3). Scale bar, 40 µm. (M–O) fERG recordings showing representative Scoptoic 3.0 waveforms (M) and quantification of a- and b-wave amplitudes (N and O). A stratified statistical analysis was performed to specifically assess gene therapy efficacy. Scale bar, 100 µV, 25 ms. Groups: Prom1 +/+ (n=3), Prom1 +/- (n=3), Prom1 -/- (n=7), and Prom1 -/- +AAV8-hPROM1 (n=8). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001 .

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Gene Knockout, Staining, Injection, Western Blot, Expressing, Labeling, Immunostaining, Infection

(A) SD-OCT images of 2-month-old mouse retinas. (B) Supplementary to . Statistical analysis of retinal thickness changes from 2 weeks to 8 months in Prom1 +/+ and Prom1 -/- mice. (C) H&E staining reveals retinal degeneration in a 1-year-old Prom1 -/- mice. Scale bar, 40 µm. (D) Retinal morphology of Prom1 +/− mice at 2 weeks, 6 weeks, and 8 months. Scale bar, 40 µm. (E) Comparison of ONL and IS/OS thickness in 6-week-old Prom1 +/+ , Prom1 +/- , and Prom1 -/- mouse retinas. (F) PROM1-N staining in Prom1 +/+ retinas. Scale bar, 50 µm. (G–I) Scotopic 3.0 recordings in PM1 mice. Panel (G) displays representative waveforms. The a- and b-wave amplitudes are quantified in (H) and (I). Groups: Prom1 +/+ (n=5), Prom1 +/- (n=6), and Prom1 -/- (n=16). (J) Extinguished fERG response in 1-year-old prom1 -/- mice. (K–M) Representative waveforms in PM2 mice (J). The amplitudes of the a- and b-wave are detailed in (K) and (L). Prom1 +/+ (n=5), Prom1 +/- (n=6), Prom1 -/- (n=7), and Prom1 -/- +AAV8-hPROM1 (n=8). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); **P < 0.01; ***P < 0.001 .

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A) SD-OCT images of 2-month-old mouse retinas. (B) Supplementary to . Statistical analysis of retinal thickness changes from 2 weeks to 8 months in Prom1 +/+ and Prom1 -/- mice. (C) H&E staining reveals retinal degeneration in a 1-year-old Prom1 -/- mice. Scale bar, 40 µm. (D) Retinal morphology of Prom1 +/− mice at 2 weeks, 6 weeks, and 8 months. Scale bar, 40 µm. (E) Comparison of ONL and IS/OS thickness in 6-week-old Prom1 +/+ , Prom1 +/- , and Prom1 -/- mouse retinas. (F) PROM1-N staining in Prom1 +/+ retinas. Scale bar, 50 µm. (G–I) Scotopic 3.0 recordings in PM1 mice. Panel (G) displays representative waveforms. The a- and b-wave amplitudes are quantified in (H) and (I). Groups: Prom1 +/+ (n=5), Prom1 +/- (n=6), and Prom1 -/- (n=16). (J) Extinguished fERG response in 1-year-old prom1 -/- mice. (K–M) Representative waveforms in PM2 mice (J). The amplitudes of the a- and b-wave are detailed in (K) and (L). Prom1 +/+ (n=5), Prom1 +/- (n=6), Prom1 -/- (n=7), and Prom1 -/- +AAV8-hPROM1 (n=8). Error bars represent mean ± S.E.M.; ns, not significant (P ≥ 0.05); **P < 0.01; ***P < 0.001 .

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Staining, Comparison

(A and B) Western blots of PROM1 in mouse retinas, showing original image (A) and technical replicate (B). β-Actin as loading control.

Journal: bioRxiv

Article Title: Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Mouse Models

doi: 10.1101/2025.08.06.668899

Figure Lengend Snippet: (A and B) Western blots of PROM1 in mouse retinas, showing original image (A) and technical replicate (B). β-Actin as loading control.

Article Snippet: A 2057 bp genomic deletion encompassing exons 2–20 of the Prom1 gene (NM_001163582) was introduced into C57BL/6JCya mouse via CRISPR/Cas9 (Cyagen, Guangzhou).

Techniques: Western Blot, Control

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