Txnip deletions and missense alleles prolong the survival of cones in a retinitis pigmentosa mouse model
- Departments of Genetics and Ophthalmology, Blavatnik Institute, Harvard Medical School, United States
- Lingang Laboratory, China
- School of Life Science and Technology, ShanghaiTech University, China
- Howard Hughes Medical Institute, United States
Figures
Figure 1
Effect of arrestin domain containing protein 4 (Arrdc4) on cone survival in retinitis pigmentosa mice.
(A) Amino acid sequences of mouse TXNIP and mouse ARRDC4. In the full-length alignment (421 amino acid), Identity: 172/421, 40.86%; Similarity: 246/421, 58.43%; Gaps: 28/421, 6.65%. Color code: …
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Figure 1—source data 1
This file contains the source data of Figure 1D.
- https://cdn.elifesciences.org/articles/90749/elife-90749-fig1-data1-v1.xlsx
Figure 2 with 2 supplements
Txnip deletions expressed only within retinal pigmented epithelium (RPE) cells: effects on GLUT1 removal and cone survival.
(A) Glucose transporter 1 (GLUT1) expression in P20 wild-type eyes infected with control (AAV8-RedO-H2BGFP, 2.5×108 vg/eye), or a Txnip allele (2.5×108 vg/eye) plus RedO-H2BGFP (2.5×108 vg/eye), as …
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Figure 2—source data 1
This file contains the source data of Figure 2C, E and G and Figure 2—figure supplement 1.
- https://cdn.elifesciences.org/articles/90749/elife-90749-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Txnip deletions expressed only within retinal pigmented epithelium (RPE) cells: quantification of the Glucose transporter 1 (GLUT1) level within the basal surface of the RPE.
GLUT1 expression in P20 wild-type (wt) eyes infected with control (AAV8-RedO-H2BGFP, 2.5×108 vg/eye), or a Txnip allele (2.5×108 vg/eye) plus RedO-H2BGFP (2.5×108 vg/eye), as indicated in each …
Figure 2—figure supplement 2
Predicted protein-protein interactions of TXNIP and Glucose transporter 1 (GLUT1) by an algorithm, ColabFold, based on AlphaFold-2.
Figure 3
Tests of Txnip alleles on cone survival.
(A) Representative P50 rd1 flat-mounted retinas after P0 infection with 1 of 5 different Txnip alleles (AAV8-RedO- N.Txnip /C.Txnip.C247S/ nt.Txnip.C247S1-301/nt.Txnip.C247S1-320 or …
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Figure 3—source data 1
This file contains the source data of Figure 3B and C.
- https://cdn.elifesciences.org/articles/90749/elife-90749-fig3-data1-v1.xlsx
Figure 4
Summary of various alleles of Txnip in this and previous study (Xue et al., 2021).
‘Retinal pigmented epithelium (RPE) Glucose transporter 1 (GLUT1) Removal’ refers to the amount of GLUT1 immunohistochemical signal on the basal surface following expression in the RPE using the …
Figure 5 with 2 supplements
Effect of knockdown of Hsp90ab1 in retinitis pigmentosa cones in vivo.
(A) AAV8-RO1.7-Hsp90ab1-FLAG (1×109 vg/eye) co-injected with shNC (non-targeting shRNA control, AAV8-RedO-shRNA, 1×109 vg/eye) or co-injected with Hsp90ab1 shRNAs #a, #b, #c (AAV8-RedO-shRNA, 1×109 …
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Figure 5—source data 1
This file contains the source data of Figure 5B, D, F and H.
- https://cdn.elifesciences.org/articles/90749/elife-90749-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Predicted 3D protein structures of HSP90AB1 and PARP1.
(A) Predicted 3D protein structures of HSP90AB1 by AI algorithm AlphaFold-2 from two angles of view. (B) Predicted 3D protein structures of PARP1 by AI algorithm AlphaFold-2 from two angles of view.
Figure 5—figure supplement 2
Predicted 3D protein interactions among TXNIP, HSP90AB1, and PARP1 by AI algorithm AlphaFold Multimer from two angles of view.
Abbreviations: Arr C-, C-terminal arrestin domain.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | GLUT1 (rabbit monoclonal) | Abcam | ab115730 | IHC (1:500) |
| Genetic reagent (M. musculus) | Arrdc4 cDNA | GeneCopoeia | Cat. #: Mm26972 NCBI: NM_001042592.2 | |
| Genetic reagent (M. musculus) | Hsp90ab1 cDNA | GeneCopoeia | Cat. #: Mm03161 NCBI: NM_008302.3 | |
| Software, algorithm | Protein 3D structure prediction | AlphaFold-2 | TXNIP (M. musculus); ARRDC4 (M. musculus); HSP90AB1 (M. musculus); PARP1 (M. musculus) | Jumper et al., 2021; https://alphafold.ebi.ac.uk |
| Software, algorithm | Protein 3D interaction prediction | ColabFold | AlphaFold2_mmseqs2 | Mirdita et al., 2022; Ovchinnikov, 2021; https://github.com/sokrypton/colabfold |
| Software, algorithm | Protein 3D interaction prediction | COSMIC2 | AlphaFold2 – Multimer | Evans et al., 2021; http://cosmic-cryoem.org/tools/alphafoldmultimer/ |
| Software, algorithm | Protein 3D structure viewer | RCSB PDB | Mol* 3D Viewer | To visualize the 3D structure of proteins in.pdb files https://www.rcsb.org/3d-view |
