jc 1 probe  (Beyotime)

Journal: Genes & Diseases

Article Title: PDK1 elevation was induced by epigenetic modifications of KDM3A and METTL16 to mediate TKI resistance and cancer development

doi: 10.1016/j.gendis.2025.101947

Figure Lengend Snippet: PDK1 inhibitor JX06 and gefitinib synergistically induced cell apoptosis in gefitinib-resistant lung cancer cells. (A) The protein expression levels of PDK were reduced upon the treatment of PDK1 inhibitor JX06 in PC-9 and PC-9/G cells. (B) B2B and PC-9/G cells were treated with different concentrations of JX06 for 48 h. The cell viabilities were determined by CCK-8. (C – E) The synergy effect between JX06 and gefitinib was determined and analyzed with CompuSyn software. (F) The apoptosis rates were analyzed with flow cytometry after the combined treatment of gefitinib and JX06. (G) The TUNEL assay was performed with the indicated treatment in PC-9/G cells. (H) The cells treated as described were stained with the JC-1 probe and detected using a fluorescence microscope. Red fluorescence indicates the aggregation form of JC-1, showing increased mitochondrial membrane potential (ΔΨm). Green fluorescence indicates the monomeric form of JC-1, which indicates reduced mitochondrial membrane potential (ΔΨm). Data were statistically analyzed with Student’s t -test, and values were shown as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: After different treatments for 48 h, the mitochondrial membrane potential of cells was detected with the enhanced mitochondrial membrane potential assay kit using JC-1 probe (Beyotime, Jiangsu, China), and the fluorescence was analyzed with a fluorescence microscope.

Techniques: Expressing, CCK-8 Assay, Software, Flow Cytometry, TUNEL Assay, Staining, Fluorescence, Microscopy, Membrane, Standard Deviation

Journal: Research

Article Title: Extracellular Vesicle-Mediated Delivery of Mitochondrial Circular RNA MTCO2 Protects against Cerebral Ischemia by Modulating mPTP-Dependent Ferroptosis

doi: 10.34133/research.1232

Figure Lengend Snippet: Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Mitochondrial transmembrane potential was assessed using the lipophilic cation probe JC-1 (MedChemExpress, USA), which accumulates in a potential-dependent manner.

Techniques: Permeability, Fluorescence, Control, Flow Cytometry, Staining, Membrane

Journal: Research

Article Title: Extracellular Vesicle-Mediated Delivery of Mitochondrial Circular RNA MTCO2 Protects against Cerebral Ischemia by Modulating mPTP-Dependent Ferroptosis

doi: 10.34133/research.1232

Figure Lengend Snippet: Mitochondria-targeted circMTCO2 delivery alleviates oxygen-glucose deprivation (OGD)-induced neuronal ferroptosis. (A) Representative fluorescence images of calcein, MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in Neuro2a (N2a) cells under control or OGD conditions, treated with phosphate-buffered saline (PBS), EV Ctrl , RVG-EV RNA , and RVG-EV mt-RNA . (B) Quantification of fluorescence intensities shown in panel (A). (C) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). (D) Cell viability assessed by the Cell Counting Kit-8 (CCK-8) assay. Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test and one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Mitochondrial transmembrane potential was assessed using the lipophilic cation probe JC-1 (MedChemExpress, USA), which accumulates in a potential-dependent manner.

Techniques: Fluorescence, Control, Saline, Flow Cytometry, Staining, Membrane, Cell Counting, CCK-8 Assay