PEDF Deletion Induces Senescence and Defects in Phagocytosis in the RPE
Abstract
:1. Introduction
2. Results
2.1. Serpinf1 Deletion Increased Senescence-Associated Gene Expression
2.2. Serpinf1 Deletion Increased Β-Galactosidase Activity in the RPE
2.3. Serpinf1 Deletion Increased the Size of Nuclei of the RPE Cells
2.4. Serpinf1 Ablation Downregulated Pnpla2 in the RPE
2.5. Rhodopsin and Lipid Accumulated in the Serpinf1-Depleted RPE
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. RPE/Choroid Flat Mounts
4.3. Immunofluorescence and Fluorescent Confocal Microscopy
4.4. Fluorescent Imaging Quantification
4.5. Transcript Levels Determination by Quantitative PCR (qPCR)
4.6. Western Blot
4.7. β-Galactosidase Activity Assay
4.8. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Protein | Forward Primer | Reverse Primer |
---|---|---|---|
Rplp0 | RPLP0 | 5′-CTTCATTGTGGGAGCAGACA-3′ | 5′-GTGAGGTCCTCCTTGGTGAA-3′ |
Serpinf1 | PEDF | 5′-ACCGTGACCCAGAACTTGAC-3′ | 5′-CACGGGTTTGCCAGTAATCT-3′ |
Pnpla2 | PEDF-R | 5′-ACAGTGTCCCCATTCTCAGG-3′ | 5′-TTGGTTCAGTAGGCCATTCC-3′ |
H2ax | H2AX | 5′-GCCTCATACCAGTTGACCCTG-3′ | 5′-TAGAACTCTTGTCCACAGGCC-3′ |
Trp53 | P53 | 5′-GACCCTGGCACCTACAATGAA-3′ | 5′-GGGGTGGATAAATGCAGACA-3′ |
Cdkn1a | P21 | 5′-ATCTGCTGCTCTTTTCCCCC-3′ | 5′-CCCTAGACCCACAATGCAGG-3′ |
Glb1 | GLB1 | 5′-GCACGGCATCTATAATGTCACC-3′ | 5′-AGCCGGTCCTCCCAGTAG-3′ |
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Rebustini, I.T.; Crawford, S.E.; Becerra, S.P. PEDF Deletion Induces Senescence and Defects in Phagocytosis in the RPE. Int. J. Mol. Sci. 2022, 23, 7745. https://doi.org/10.3390/ijms23147745
Rebustini IT, Crawford SE, Becerra SP. PEDF Deletion Induces Senescence and Defects in Phagocytosis in the RPE. International Journal of Molecular Sciences. 2022; 23(14):7745. https://doi.org/10.3390/ijms23147745
Chicago/Turabian StyleRebustini, Ivan T., Susan E. Crawford, and S. Patricia Becerra. 2022. "PEDF Deletion Induces Senescence and Defects in Phagocytosis in the RPE" International Journal of Molecular Sciences 23, no. 14: 7745. https://doi.org/10.3390/ijms23147745