Journal: Communications Biology

Article Title: Targeting Cend1-Atp5f1b interaction rescues mitochondrial dysfunction and ameliorates ischemic brain injury

doi: 10.1038/s42003-025-09419-4

Figure Lengend Snippet: 293 T cells were transfected with empty vector, wild-type Cend1, or the Cend1-G130P mutant for 48 h, followed by oxygen–glucose deprivation/reoxygenation (OGD/R) treatment. Cells were collected 24 h after reoxygenation for subsequent analyses. A Representative flow cytometric analysis of JC-1 staining showing the proportion of monomers and aggregates. B Quantification of JC-1 monomer proportion. N = 3. C Representative flow cytometric analysis of calcein–cobalt staining to assess mPTP opening. D Quantification of calcein fluorescence intensity. N = 3. E Relative NADH–CoQ reductase (complex I) activity measured using a mitochondrial complex I assay kit. N = 3. F Relative F₁F₀-ATP synthase (complex V) activity measured using a mitochondrial complex V assay kit. N = 3. G Intracellular ATP levels measured by ATP assay kit and normalized to protein content. N = 3. Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; ns, not significant.

Article Snippet: Activities of mitochondrial complex I and V in 293 T cells were measured using the Cell Mitochondrial Complex I Activity Assay Kit (Elabscience, #E-BC-K834-M) and Complex V Activity Assay Kit (Elabscience, #E-BC-K838-M), respectively, according to the manufacturer’s instructions.

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Staining, Fluorescence, Activity Assay, ATP Assay

Journal: Communications Biology

Article Title: Targeting Cend1-Atp5f1b interaction rescues mitochondrial dysfunction and ameliorates ischemic brain injury

doi: 10.1038/s42003-025-09419-4

Figure Lengend Snippet: Cells were collected 24 h after reperfusion for subsequent assays. A Cells were stained with JC-1 at working concentration, and the ratio of JC-1 monomers to aggregates was analyzed by flow cytometry. B Quantification of JC-1 monomer proportion. N = 3. C Cells were stained with MPTP detection reagent at working concentration, and green fluorescence intensity was analyzed by flow cytometry. D Quantification of green fluorescence intensity reflecting mPTP opening. N = 3. E NADH–CoQ reductase activity was measured using a mitochondrial complex I activity assay kit according to the manufacturer’s instructions, and relative activity was quantified. N = 3. F F₁F₀-ATP synthase activity was measured using a complex V activity assay kit and quantified as relative activity. N = 3. G Intracellular ATP content was measured using a luminescence-based ATP assay kit and quantified relative to control. N = 3. Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; ns, not significant.

Article Snippet: Activities of mitochondrial complex I and V in 293 T cells were measured using the Cell Mitochondrial Complex I Activity Assay Kit (Elabscience, #E-BC-K834-M) and Complex V Activity Assay Kit (Elabscience, #E-BC-K838-M), respectively, according to the manufacturer’s instructions.

Techniques: Staining, Concentration Assay, Flow Cytometry, Fluorescence, Activity Assay, ATP Assay, Control