Fig. 4: Leptin provoked fused mitochondrial network under GSDH condition.
a Mitochondrial ultrastructures were analyzed by electron micrograph with representative images showing significant changes in mitochondrial length after hMSCs were pretreated with leptin (50 ng/ml) as compared with those pretreated with the solvent alone (magnification was set at × 15,000 and × 50,000, respectively); scale bar, 1 μm. b The mitochondrial length was measured using Adobe Photoshop CS5 for at least 20 mitochondria for each cell (at least 30 cells for each group). c OCR was quantified for both hMSCs-Leppre and hMSCs-Ctrlpre using OROBOROS instrument. d Cellular ATP levels were conducted through luciferin/luciferase-based assay, and the data were calibrated with protein content. e, f Protein expression implicated mitochondrial homeostasis including fusion, fission, and biogenesis, and was assessed by western blot for hMSCs-Leppre and hMSCs-Ctrlpre and quantified by densitometry using β-actin as loading control. g Expression levels of genes involved in mitochondrial homeostasis were detected with qRT-PCR normalized to that of β-actin. h The differential expression levels of L-OPA1 and S-OPA1 isoforms can be identified by western blot using the specific antibody for hMSCs-Leppre or hMSCs-Ctrlpre in GSDH conditions for 24 h. Each experiment was repeated three times. All data were shown as mean ± SEM. *denotes P < 0.05, **P < 0.01
