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Isolation of PDGFRβ+ REPs

GG Guangfeng Geng
QC Quan Chen
YZ Yushan Zhu
LS Lihong Shi

Last updated date: Aug 15, 2024 Views: 101 Forks: 0

original An abbreviated version of this protocol was published in eLife in May, 2021
Receptor-mediated mitophagy regulates EPO production and protects against renal anemia
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Sorting of mouse PDGFRβ+ cells

 

  1. Harvest a mouse kidney and remove the fibrous capsule.
  2. Kidney was minced with scissors at room temperature in RPMI 1640.
  3. Gently mix the minced kidney with RPMI 1640 and transfer into tubes.
  4. Incubated the minced kidney in RPMI 1640 medium containing 0.5 mg/mL Liberase(Cat# 423610, Roche) and 100 U/mL DNase I (Cat# D8071, Solarbio) at 37°C for at least 45 min.
  5. Add 0.1% bovine serum albumin to stop the enzymatic reaction.
  6. After filtration and red blood cell lysis, Centrifuge the mixture for 10 min and remove the supernatant.
  7. The remaining single cells were incubated with antibody(Anti-mouse PDGFR beta Antibody, Clone APB5) before BD FACSAria™ III flow cytometry analysis.
Copyright: © 2024 The Author(s); This is an open-access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Geng, G, Chen, Q, Zhu, Y and Shi, L(2024). Isolation of PDGFRβ+ REPs. Bio-protocol Preprint. bio-protocol.org/prep2713.
  2. Geng, G., Liu, J., Xu, C., Pei, Y., Chen, L., Mu, C., Wang, D., Gao, J., Li, Y., Liang, J., Zhao, T., Zhang, C., Zhou, J., Chen, Q., Zhu, Y. and Shi, L.(2021). Receptor-mediated mitophagy regulates EPO production and protects against renal anemia. eLife. DOI: 10.7554/eLife.64480

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