Journal: Biology

Article Title: Mitochondrial Ultrastructure and Activity Are Differentially Regulated by Glycolysis-, Krebs Cycle-, and Microbiota-Derived Metabolites in Monocytes

doi: 10.3390/biology11081132

Figure Lengend Snippet: Lactate, succinate, fumarate, butyrate, and acetate induce mitochondria polarization in monocytes. Mitochondrial membrane potential (Δψ m ) was assessed in human monocytes by flow cytometry, using the tetramethylrhodamine methyl ester molecular probe. For analysis, CD14+ cells were gated and TMRM mean fluorescence intensity (indicative of Δψ m ) was recorded. ( A ) shows the Δψ m fold change at 1, 2, 3, and 4 h post metabolite exposure, ( B ) shows representative histograms of TMRM fluorescence. Results are from eight independent experiments. Mitochondrial polarization was statistically significant ( p < 0.001, Kruskal–Wallis and Dunn’s post hoc tests).

Article Snippet: PBMC were subjected to several low-speed washes in PBS in order to minimise platelet contamination and were subsequently subjected to monocyte enrichment by negative selection using a monocyte-enrichment cocktail and magnetic separation on an EasySep magnet or RoboSep instrument (Stem Cell Technologies, Vancouver, BC, Canada) or by positive selection using CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Membrane, Flow Cytometry, Fluorescence