Fig. 7: Efferocytosis shifts MSC mitochondria toward fission. | Cell Death & Disease

Fig. 7: Efferocytosis shifts MSC mitochondria toward fission.

From: Efferocytosis by bone marrow mesenchymal stromal cells disrupts osteoblastic differentiation via mitochondrial remodeling

Fig. 7

A Heat map of mitochondrial biogenesis and dynamic genes and pathways at 3 h or 24 h vs the control in ST2 cells. Key genes regulating mitochondrial dynamics are highlighted in red (fission) and blue (fusion). B, C Representative images of control and efferocytically challenged (PMN+) hMSCs stained with MitoTracker Red (mitochondria) and DAPI (nucleus). PMN stained with Calcein AM. Highlighted regions are magnified and shown as inserts on the right of each image. ImageJ analysis of mitochondrial connectivity shown as numbers of connected pixels (Mean ± SD, n = 25). D Mitochondria from control and challenged (PMN+) hMSCs stained with tetramethyl rhodamine ethyl ester (TMRE) to measure mitochondrial membrane potential. Graphs show the biological data points N = 3, Mean + SD, *p < 0.05, (t-test). E Quantification of efferocytosis by hMSCs incubated excess (1:10) end stage neutrophils (hPMNs) over 24 h. N = 4 at each time point. Means ± SD are shown. (F) Representative microscopy images showing uptake of end stage neutrophil (GFP+) by hMSC (mCherry+) and the resulting voids in the cytoplasm (white arrows). G Experimental Schematic for samples treated with a mitochondrial inhibitor (MDivi). H–J Quantification via flow cytometry of viability, end stage neutrophil (PMNs) engulfment, and efficiency (mean fluorescent intensity, MFI) of hMSCs pre-treated with 25 μM Mdivi for 1 h and then given end stage PMN for 24 h. K Quantification of Alkaline Phosphatase (ALP) via qPCR of hMSCs pre-treated with 25 μM Mdivi for 1 h and then given end stage PMN for 3 h. N = 3, Mean ± SD shown on graphs **p < 0.01, ***p < 0.001, (t-test or ANOVA).

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