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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
JoVE Journal
Biology
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JoVE Journal Biology
Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
DOI:

15:43 min

January 07, 2013

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Chapters

  • 00:05Title
  • 02:27Primary Cultures of Renal Proximal Tubular Cells
  • 05:24Measurement of RPTC Respiration
  • 06:18Analysis of Mitochondrial Membrane Potential (ΔΨm)
  • 07:14Isolation of Mitochondria
  • 08:16Assaying Enzyme Complexes of the Respiratory Chain
  • 10:24Measurement of Intracellular ATP Content
  • 10:55Analysis of Cell Viability
  • 12:18Results: Overexpressing PKC-ε in RPTCs
  • 14:56Conclusion

Summary

Automatic Translation

The effects of activation of protein kinase C (PKC) isozymes on mitochondrial functions associated with respiration and oxidative phosphorylation and on cell viability are described. The approach adapts adenoviral technique to selectively overexpress PKC isozymes in primary cell culture and a variety of assays to determine mitochondrial functions and energy status of the cell.

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