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

Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes

DOI:
10.3791/4301-v

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15:43 min

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January 07, 2013

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1Department of Pharmaceutical Sciences, College of Pharmacy,University of Arkansas for Medical Sciences

Chapitres

  • 00:05Titre
  • 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

Traduction automatique

Traduction automatique

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.

Tags

Protein Kinase CPKC IsozymesMitochondrial FunctionCellular Energy StatusCardioprotectionNephrotoxicant-induced Mitochondrial DysfunctionCell DeathRenal CellsOxidative PhosphorylationCell SurvivalRenal Proximal Tubular Cells (RPTC)Adenoviral VectorsPKC-epsilon Mutant CDNAs

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