High throughput respiration assay on a 96 well format (cell lines or tissue) :

Different energy states are measured :

• Basal respiration
• Non-phosphorylating respiration (state 4)
• Phosphorylating respiration (state 3)
• Uncoupled respiration (maximal)

Different energy substrates can be used

Oxygen kinetics ( high resolution respirometry ; P50 determination)

Energy transduction assays (Cell lines or tissue) :

• ATP content measurement (total ATP, mitochondrial ATP, glycolytic ATP, steady state and flux)
• Ratio ADP/ATP
• Energy safety margin and biochemical threshold effect determination

Mitochondrial membrane electric potential

Respiratory chain complexes activity (Complex I, II, III, IV, I+III, II+III, I+IV, II+IV enzyme activity)

Oxidative stress and antioxidant activities :

• Reduced glutathione (GSH) / oxidized glutathione (GSSG) Ratio
• Intracellular ROS levels (total, mitochondrial)
• Oxiblots (protein damages)
• Redox statuts: NADH/NAD+
• Western Blots on key REDOX regulators

Mitochondrial content (Mitochondrial mass (mtDNA/nDNA) and turnover (biogenesis/mitophagy))

Mitochondrial dynamics :

• Network morphology and distribution; fission assay
• Immunocytochemistry on selected markers involved in fusion, fission, motility
• Western Blot analysis of fusion and fission proteins (expression and posttranscriptional modification)

Energy sensing and signaling pathway analysis (Westernblot on metabolic reprogramming master regulators and effectors)

Energy substrates consumption assays during cell culture

Cell proliferation assay on different substrates (metabolic addiction studies)

Proteomics (the analysis can be performed on cell lines or tissues (human, mouse….) :

• PAGE- Analysis LC* MS/MS LTQ Orbitrap Q exactive
• Bioinformatic analysis of the metabolic changes :
  • IPA analysis (Ingenuity pathway)
  • KEGG pathway analysis
  • Canonical pathway analysis
  • CELLOMET non* canonical pathway analysis (situations of metabolic remodeling)
  • List of the molecular and cellular functions
  • List of physiological system Development and function
  • List of top upstream regulators
  • List of biomarkers and their active compounds
  • List of diseases and functions
  • Metabolic signature

Western Blot on selected key metabolic proteins (according to the project)

Transcriptomics :

• Targeted (custom qPCR plates)
• Genomic wide transcriptomics

• Identification and validation of biomarkers from the analysis of omic data (proteomic, transcriptomic) obtained from patient sample
• Validation by WesternBlot and qPCR
• Analysis of omic data (proteomic, transcriptomic) obtained from patient samples

• Academic research and projects (grants…)
• Lectures (training course)
• R&D industry metabolism-oriented programs