mitochondrial membrane potential δψm  (TargetMol)

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR attenuate oxidative stress, preserve mitochondrial integrity, and suppress senescence in TSPCs by inhibiting the mtDNA-STING-NF-κB axis. (A) Schematic of the experimental design. (B) TSPC proliferation assessed by CCK-8 assay. (C, E) Immunofluorescence staining and quantification of HSP70 (n = 3). (D, F) Mitochondrial membrane potential (ΔΨm) visualized by JC-1 staining (red: high potential; green: low potential) and quantification. (G) Multi-SIM of mtDNA (magenta) and TOMM20 (green) with colocalization analysis. (H) Western blot of cGAS-STING-IRF3-NF-κB pathway proteins. (I, K) Colony-forming unit fibroblast (CFU-F) assay and quantification of self-renewal capacity (n = 3). (J) Senescence-associated β-galactosidase (SA-β-gal) activity. (L, M) Apoptosis analysis by Annexin V/PI flow cytometry and quantification (n = 3). Scale bars: 100 μm (C); 5 μm (D, G); 200 μm (J). Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Mitochondrial membrane potential (ΔΨm) was evaluated via JC-1 staining (MedChemExpress, USA).

Techniques: CCK-8 Assay, Immunofluorescence, Staining, Membrane, Western Blot, Activity Assay, Flow Cytometry

Journal: Journal of Advanced Research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: P2rx7 controls mitochondrial dynamics in MSCs. a OCR, calculated basal respiration and ATP-linked respiration of the control and the P2rx7 -/- MSCs. b ATP contents of the control and the P2rx7 -/- MSCs. c ECAR, and calculated glycolysis, glycolytic capability and glycolytic reverse of the control and the P2rx7 -/- MSCs. d TEM images of mitochondria (arrows) in the control and the P2rx7 -/- MSCs (n = 20 per group). e ΔΨM of the control and the P2rx7 -/- MSCs was measured by JC-1 staining (polymer: red; monomer: green). f The expressions of genes Nd1 and mt-Cytb in the control and the P2rx7 -/- MSCs that related to the mitochondrial mass in cells assessed by qPCR. g ΔΨM of MSCs treated with different concentrations of BzATP was analyzed by JC-1 staining. Scale bar: d: 500 nm; e, g: 100 μm. Data was presented as mean ± SD. The data (a, b, e-g) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (a-f) and one-way ANOVA (g). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Mitochondrial membrane potential was evaluated by Mitochondrial membrane potential (ΔΨM) assay kit with JC-1 (Beyotime, Shanghai, China) in accordance to the manufacturer’s manual.

Techniques: Control, Staining, Polymer, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: P2rx7 enhanced the sensitivity of ERK pathway to promote mitochondrial fusion. a Top 10 significantly enrichment of downregulated DEGs between control and P2rx7 -/- MSCs analyzed by KEGG. b Expressions of Erk1/2 and p-Erk1/2 protein in the distal femurs were analyzed by IHC staining (n = 5 per group). c The phosphorylation level and the total expression level of Erk1/2 in the control and the P2rx7 -/- MSCs before and after the activated by bFGF, as assessed by western blotting. d The phosphorylation level and the total expression level of Erk1/2 in MSCs treated with different concentrations of BzATP before and after the activated by 10 μM bFGF for 1 h, as assessed by western blotting. e Expressions of genes associated with mitochondrial dynamics in the Widetype (WT) and the P2rx7 -/- MSCs treated with bFGF as assessed by western blotting. f Expressions of genes associated with mitochondrial dynamics in MSCs treated with siNC, siMfn1 and BzATP as assessed by western blotting. g ΔΨM of the Widetype (WT) and the P2rx7 -/- MSCs treated with BzATP and bFGF was analyzed by JC-1 staining. h ALP activity (top) and mineralized nodules (bottom) under osteogenic induction of human MSCs when exposed to 100 nM BzATP in the presence and absence of U0126. i The expression of osteogenesis marker genes in MSCs exposed with BzATP, in the presence and absence of 10 μM U0126. Scale bar: b: 100 μm; d: 50 μm; h: 500 μm. Data was presented as mean ± SD. The data (c-i) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (b) and one-way ANOVA (g, h).

Article Snippet: Mitochondrial membrane potential was evaluated by Mitochondrial membrane potential (ΔΨM) assay kit with JC-1 (Beyotime, Shanghai, China) in accordance to the manufacturer’s manual.

Techniques: Control, Immunohistochemistry, Phospho-proteomics, Expressing, Western Blot, Staining, Activity Assay, Marker, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: Multi-omics analysis reveals a protective role of endogenous proline in sheep oocyte vitrification and its therapeutic application

doi: 10.3389/fcell.2025.1714529

Figure Lengend Snippet: Impact of L-proline on mitochondrial function in vitrified oocytes. (A) Mitochondrial superoxide detection via MitoSOX™ Red staining. Higher fluorescence intensity indicates elevated mitochondrial ROS levels. Representative micrographs are shown: fresh, vitrified, and vitrified + L-P. Scale bar: 100 μm. (B) Mean fluorescence intensity of MitoSOX™ Red across the experimental groups. (C) The CM potential was detected by JC-1 staining. At high mitochondrial membrane potential (ΔΨm), JC-1 accumulates in the mitochondrial matrix and forms J-aggregates that emit red fluorescence (590 nm), indicative of functional mitochondria. Conversely, under low ΔΨm, JC-1 remains monomeric throughout the cytoplasm, emitting green fluorescence (530 nm), indicating mitochondrial depolarization and dysfunction. (D) JC-1 fluorescence ratio (red:green) across experimental groups, histogram representing the ratio of red fluorescence (functional mitochondria) to green fluorescence (depolarized mitochondria) in the oocyte groups. (E) ATP quantification via the BODIPY™ FL ATP probe. The fluorescence intensity is directly proportional to the ATP concentration. Scale bar: 50 μm. (F) Mean fluorescence intensity of BODIPY™ FL ATP across experimental groups. The data are expressed as the means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns represents no significance.

Article Snippet: Mitochondrial Membrane Potential (ΔΨm) was assessed using a JC-1 assay kit according to the manufacturer’s instructions (Solarbio, #CA1310).

Techniques: Staining, Fluorescence, Membrane, Functional Assay, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Multi-omics analysis reveals a protective role of endogenous proline in sheep oocyte vitrification and its therapeutic application

doi: 10.3389/fcell.2025.1714529

Figure Lengend Snippet: Regulatory effects of L-proline on calcium homeostasis in vitrified Ovine Oocytes. (A) Representative Ca 2+ fluorescence images across experimental groups. (B) Quantitative analysis of the mitochondrial Ca 2+ (Rhod-2 a.m.), endoplasmic reticulum Ca 2+ (Mag-Fluo-4 a.m.), and cytosolic Ca 2+ (Fluo-3 a.m.) levels. The data are expressed as the means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns represents no significance.

Article Snippet: Mitochondrial Membrane Potential (ΔΨm) was assessed using a JC-1 assay kit according to the manufacturer’s instructions (Solarbio, #CA1310).

Techniques: Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Multi-omics analysis reveals a protective role of endogenous proline in sheep oocyte vitrification and its therapeutic application

doi: 10.3389/fcell.2025.1714529

Figure Lengend Snippet: (A) After cryopreservation, the endogenous synthesis of osmoprotectants such as proline, taurine, and inositol in ovine oocytes is increased to counteract osmotic stress and low-temperature changes. Supplementation of ovine oocyte freezing medium with 0.5 M L-proline significantly improved the survival rate of cryopreserved oocytes. (B) It reduces oxidative stress induced by freezing, ameliorates the distribution of organelles (including the spindle apparatus, endoplasmic reticulum, and mitochondria), lowers mitochondrial reactive oxygen species (ROS) levels, increases the mitochondrial membrane potential, mitigates the decline in ATP levels caused by cryopreservation, and maintains intracellular calcium homeostasis.

Article Snippet: Mitochondrial Membrane Potential (ΔΨm) was assessed using a JC-1 assay kit according to the manufacturer’s instructions (Solarbio, #CA1310).

Techniques: Membrane