Adenosine A2b receptor antagonist PSB1115 was purchased from R&D Systems and dissolved in PBS (pH adjusted to 7.4). As described before (He et al., 2014 (link)), 10 mg/kg PSB1115 was injected via the tail vein 20 min before ischemia. Vehicle groups were given the equivalent vehicle solution as a control.
Psb1115
Manufactured by R&D Systems
The PSB1115 is a laboratory equipment product. It is a device designed for use in research and development settings. The core function of the PSB1115 is to perform a specific task or set of tasks related to the research and analysis needs of the laboratory.
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3 protocols using psb1115
1
PSB1115 Antagonist Ischemia Pretreatment
2
Corneal Epithelium Regeneration in Diabetic Mice
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Mouse entire corneal epithelium, including the limbal region, was marked with 3 mm trephine and subsequently scraped with a corneal rust ring remover (Alger Co, Lago Vista, TX) in anesthetized normal and diabetic mice. After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μL/eye, R&D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control. For topical netrin-1 application, netrin-1 (50 ng in 2.5 μL PBS) was dropped onto the corneal surface by a 10 μL tip four times daily per eye for 3 days. For netrin-1 receptor inhibition, A2BAR antagonist PSB1115 (2.5 μg in 5 μL PBS, R&D Systems) or UNC5B receptor-blocking antibody (5 μg in 5 μL PBS, R&D Systems) was injected subconjunctivally at 24 h before and 0, 24, and 48 h after the scrape of corneal epithelium. Ofloxacin eye drops were applied to all mice to avoid infection. After 24, 48, and 72 h, corneal epithelium defects were visualized by staining with fluorescein sodium and photographed under slit lamp microscope (BQ900; Haag-Streit, Bern, Switzerland). The stained area on the residual epithelial defect was analyzed by using Image J software (National Institutes of Health, Bethesda, MD) with the original epithelial defect area as 100%.
3
Mitochondrial Membrane Potential in HSCs
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1,000 CD150+CD48−KSL HSCs derived from C57BL/6-Ly5.2 mice were cultured with 30,000 MPs (lineage−c-Kit+Sca-1−) or CD45+ cells obtained from BM of C57BL/6-Ly5.1 mice in S-Clone SF-03 medium supplemented with 0.5% bovine serum albumin, 0.5 ng/ml mouse SCF, and 0.5 ng/ml mouse TPO. After 48 h, the ΔΨm of HSCs was examined by staining with an antibody for CD45.2 following JC-1 staining. Before the analysis using a flow cytometer, 1 µg/ml propidium iodide (Sigma) was added into cell suspension to assess their viability. To confirm the contribution of adenosine to the suppression of ΔΨm in HSCs, 10 µM SCH442416 (Sigma), and 10 µM PSB1115 (R&D Systems) were supplied as antagonists for Adora2a and Adora2b, respectively.
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