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SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry

SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry

DOI:
10.3791/55019-v

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10:16 min

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January 06, 2017

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,
1Department of Biomedicine,Aarhus University

Chapters

  • 00:05Title
  • 00:55Western Blotting of CNTFRα Lysates
  • 03:21Immunocytochemistry in Combination with Lysosomal Inhibitors
  • 06:00Western Blot-detection of the pSTAT3 Levels
  • 08:17Results: Demonstration of CNTFRα-downregulation by Western Blotting, and Inhibition of Lysosomal Enzymes, and Immunocytochemistry
  • 09:43Conclusion

Summary

Automatic Translation

Automatic Translation

The present protocol describes how Western blotting and immunocytochemistry combined with inhibition of lysosomal enzymes can be used to demonstrate the downregulation of Ciliary Neurotrophic Factor Receptor-α (CNTFRα), which is conveyed by the interaction between Cytokine-like Factor-1 (CLF-1) and the endocytic receptor sorLA.

Tags

Keywords: SorLACLC:CLF-1CNTFRWestern BlottingImmunocytochemistryLysosomal DegradationGPI-anchored ReceptorProtein TurnoverStable Cell LinesTransfectants

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