Targeting the vascular-specific phosphatase PTPRB protects against retinal ganglion cell loss in a pre-clinical model of glaucoma
- Northwestern University Feinberg School of Medicine, United States
- Max Planck Institute for Molecular Biomedicine, Germany
Figures
Figure 1 with 2 supplements
Deletion of one Ptprb allele leads to increased TEK phosphorylation.
(A) Western blot of lung lysate from P5 Control, Tek+/-, PtprbNLS-LacZ/WT and Tek+/-;PtprbNLS-LacZ/WT mice revealed a 50% reduction in PTPRB expression in PtprbNLS-LacZ/WT (heterozygous) and Tek+/-;P…
Figure 1—figure supplement 1
Uncropped images corresponding to the western blots presented in Figure 1A.
Figure 1—figure supplement 2
Uncropped images corresponding to the western blots presented in Figure 1B.
Figure 2
TEK signaling has a dose-dependent effect on Schlemm’s canal (SC) area and development.
(A) Confocal microscopy of whole mount eyes revealed reduced CD31+ SC area in adult Tek+/- haploinsufficient mice. This phenotype was blunted in Tek+/-;PtprbNLS-LacZ/WT double heterozygous animals, …
Figure 3 with 1 supplement
Ptprb heterozygosity prevents ocular hypertension and RGC loss in Tek haploinsufficient mice.
(A) Elevated intraocular pressure (IOP) was observed in Tek+/- haploinsufficient mice at 30 weeks of age when measured by rebound tonography. As in Figure 2, this phenotype was prevented in Tek+/-;Pt…
Figure 3—figure supplement 1
Retinal vasculature was normal in Tek and Ptprb haploinsufficient mice.
(A) Confocal microscopy of P5 retinas stained with anti-CD31 antibody reveal normal progression of the superficial vasculature at both arterial and venous sprouting fronts. n = 6 (WT), 5 (Tek+/-), 3 …
Tables
Key resources table
| Resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic Reagent (M. musculus) | PtprbNLS-LacZ | Bäumer et al., 2006 | Maintained on a mixed background | |
| Genetic Reagent (M. musculus) | Tek+/- | Thomson et al., 2017 | Tektm1.1Vlcg; MGI:5544795 | Maintained on a mixed background |
| Antibody | anti-PTPRB (Rabbit polyclonal) | Nawroth et al., 2002 | Western blot: 1:2000 | |
| Antibody | Anti-TEK (Rabbit polyclonal) | Santa Cruz Biotech | sc-324 | Western blot 1:2500 |
| Antibody | anti-αTubulin (Mouse monoclonal) | Santa Cruz Biotech | sc-32293 | Western blot: 1:10,000 |
| Antibody | 4G10 Platinum anti-phosphotyrosine (Mouse monoclonal) | Millipore | 05–1050 | Western blot: 1:2000 |
| Antibody | anti-CD31 MEC13.3 (Rat monoclonal) | BD Biosciences | 55337 | IF: 1:100 |
| Antibody | anti-PROX1 (Goat polyclonal) | R and D Systems | AF2727 | IF: 1:200 |
| Antibody | anti-Ki-67 (Rabbit monoclonal) | ThermoFisher | MA5-14520 | IF: 1:200 |
| Antibody | anti-BRN3b (Goat polyclonal) | Santa Cruz Biotech | sc-6026 | IF: 1:1000 |
| Software, algorithm | ImageJ Fiji | Schindelin et al., 2012 | Version 1.52 p | Used for all image analysis |
| Software, algorithm | Graphpad Prism | Graphpad.com | Version 5.0 | Used for statistical analysis and graph generation |
| Software, algorithm | Adobe Indesign | Adobe.com | Version 14.01 × 64 | Used for figure creation |
