mitotrackertm deep red fm  (MedChemExpress)

Journal: Materials Today Bio

Article Title: ATG7-driven mitophagy in BMSC@CS hydrogel reprograms metabolism to boost bone regeneration

doi: 10.1016/j.mtbio.2025.102483

Figure Lengend Snippet: The effects of ATG7 overexpression in MSCs on mitochondrial function and energy metabolism. (A) Schematic illustration of the effects of ATG7 overexpression on mitochondrial function and energy metabolism in MSCs. (B) The mRNA levels of GLUT-3, GLUT-4, Ndufs1, Ndufs5, Ndufs6, Cyc1, CYTB and Uqcrfs1 were detected by RT‒qPCR in MSCs overexpressing ATG7. (C) ROS staining and (D) ROS FACs analysis of MSCs overexpressing ATG7. Scale bar = 75 μm. (E) TMRM staining and (F) TMRM FACs analysis of MSCs overexpressing ATG7. Scale bar = 75 μm. (G) lactate levels in the cell culture supernatant and (H) intracellular ATP levels were measured in MSCs with overexpression of ATG7.

Article Snippet: Intracellular ROS levels were assessed by incubating cells with 5 μM H 2 DCFDA (MCE, HY-D0940) in PBS for 30 min at 37 °C in the dark, followed by mounting with DAPI-containing medium (Solarbio, S2110) or staining with 1 mM mitochondrial tracker (MCE, HY-D1783) for 30 min ΔΨm was evaluated using 10 μM TMRM (MCE, HY-D0984A) under the same conditions.

Techniques: Over Expression, Staining, Cell Culture

Journal: Materials Today Bio

Article Title: ATG7-driven mitophagy in BMSC@CS hydrogel reprograms metabolism to boost bone regeneration

doi: 10.1016/j.mtbio.2025.102483

Figure Lengend Snippet: ATG7 Regulates Osteogenic Function via Mitochondrial Autophagy Activation in MSCs. (A) Western blot analysis of P62, Pink1, Parkin, and LC3 Ⅱ/Ⅰ protein expression in MSCs overexpressing Atg7. (B) Microscopy images and quantification of showing autophagy in BMSCs after overexpression ATG7. Mitochondria are stained green with Mitotracker, and lysosomes are stained red with Lysotracker. Yellow arrows indicate regions of mitochondrial and lysosomal co-localization. Scale bars: 25 μm. (C) Western blot analysis and quantification of P62, Pink1, Parkin, and LC3 Ⅱ/Ⅰ protein expression in Mdivi-1 (100 μM, 48 h) pretreated MSCs overexpressing ATG7. (D) RT‒qPCR analysis of osteogenic marker genes (ALP, OPN, OCN, and DMP-1) in ATG7-overexpressing MSCs pretreated with Mdivi-1 (100 μM) for 48 h. (E) ALP staining and quantitative analysis of osteoblast colony formation in Mdivi-1-pretreated MSCs overexpressing ATG7. Scale bar = 250 μm. (F) Western blot analysis and (G) quantification of AKT, p-AKT, PI3K, and p-PI3K protein expression in Mdivi-1 pretreated MSCs overexpressing ATG7. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Intracellular ROS levels were assessed by incubating cells with 5 μM H 2 DCFDA (MCE, HY-D0940) in PBS for 30 min at 37 °C in the dark, followed by mounting with DAPI-containing medium (Solarbio, S2110) or staining with 1 mM mitochondrial tracker (MCE, HY-D1783) for 30 min ΔΨm was evaluated using 10 μM TMRM (MCE, HY-D0984A) under the same conditions.

Techniques: Activation Assay, Western Blot, Expressing, Microscopy, Over Expression, Staining, Marker