Table 1. Molecular “Tool-Kit” for Investigating the Role of Mitochondria in Metabolic Heterogeneity & Cancer Stem Cells.

Process Interrogated	Probe or Inhibitor	Output/Assay
Mitochondrial Biogenesis	PGC1ɑ-eGFP	Metabolic Fractionation: Flow Cytometry and 3D-Spheroids
Mitochondrial ROS	Dihydro-Rhodamine 123	Metabolic Fractionation: Flow Cytometry and 3D-Spheroids
Mitochondrial H2O2	Mito-PY1	Metabolic Fractionation: Flow Cytometry and 3D-Spheroids
NADH/NADPH Levels	Auto-fluorescence (470-nm)	Metabolic Fractionation: Flow Cytometry and 3D-Spheroids
Mitochondrial Protein Synthesis	Actinonin	3D-Spheroids
NAD(+) Salvage Pathway	FK866	3D-Spheroids
Mitochondrial OXPHOS	CAPE	Seahorse XFe96 Analyzer and 3D-Spheroids
Glycolysis	2-DG, Ascorbic Acid,	3D-Spheroids Silibinin, Stiripentol