Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 3 Dysfunction of mitochondria in iPSC-derived neurons due to mutation in TIMM8A. A, B No differences in HSP60, TOMM40, and TIMM23 levels were observed among CTRL-, MUT-, and MTS-neurons by Western blot. Relative protein levels were quantified and normalized to β-Tubulin as a loading control. n = 3 independent biological replicates. C, D. Western blot analysis showed decreased levels of COX4 in mitochondrial complex IV in MUT- and MTS-neurons. Relative levels of COX4 were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. E The activity of mitochondrial complex IV was decreased in MUT- and MTS-neurons compared to CTRL-neurons. n = 3 independent biological replicates. F ATP production was decreased in MUT- and MTS-neurons compared to CTRL- neurons. n = 3 independent biological replicates. G ROS levels were increased in MUT- and MTS-neurons compared to CTRL-neurons. n = 3 independent biological replicates. H, I Levels of Cyt c and cleaved caspase-3 in CTRL-, MUT-, and MTS-neurons after treatment with 100 µM TBHP for 24 h were analyzed by Western blot. Relative protein levels of Cyt c and cleaved caspase-3 in the cytosol were quantified and normalized to β-Tubulin as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by one-way ANOVA with Tukey’s post hoc test: ns, not significant; *p < 0.05; **p < 0.01; ****p < 0.0001 (applied to panels B, D–G, and I).

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Mutagenesis, Western Blot, Control, Activity Assay

Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 4 Altered gene expression in iPSC-derived neurons revealed by RNA sequencing. A A total of 2,843 genes were significantly upregulated, and 1,667 genes were significantly downregulated in MTS-neurons compared to CTRL-neurons (adjusted p < 0.05 and |log2(fold change)| > 1). Top altered genes, including CHCHD2, are labeled. B Top 10 altered biological processes in MTS-neurons compared to CTRL-neurons, identified by Gene Ontology (GO) enrichment analysis (adjusted p < 0.05). C Top 10 altered Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in MTS-neurons compared to CTRL-neurons (adjusted p < 0.05). D Representative differentially expressed genes involved in mitochondria-related apoptosis. E Western blot analysis of the anti-apoptotic protein BCL-2 and the pro-apoptotic protein BAX in isolated mitochondrial lysates from CTRL- and MTS-neurons. F Western blot analysis of CHCHD2 in isolated mitochondrial lysates from CTRL-, MUT-, and MTS-neurons. G Expression levels of BCL-2 and BAX were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by two-tailed unpaired Student’s t test: ns, not significant; **p < 0.01. H CHCHD2 expression levels were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by one-way ANOVA with Tukey’s post hoc test: **p < 0.01; ****p < 0.0001.

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Gene Expression, Derivative Assay, RNA Sequencing, Labeling, Western Blot, Isolation, Expressing, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 6 CHCHD2 overexpression rescues mitochondrial morphology defects in iPSC-derived neurons. A, B Mitochondrial morphology in CTRL- and MTS-neurons at day 30, as revealed by transmission electron microscopy (TEM). Scale bar: 1 μm. n = 3 independent biological replicates. Quantification of mitochondrial perimeter, area, aspect ratio, and form factor was performed (103 mitochondria for CTRL group; 102 for MTS group). C, D Mitochondrial morphology in the Vector group and CHCHD2-Flag group of MTS-neurons at day 30, as revealed by TEM. Scale bar: 1 μm. n = 3 independent biological replicates. Quantification of mitochondrial perimeter, area, aspect ratio, and form factor was performed (107 mitochondria for Vector group; 124 for CHCHD2-Flag group). Data are represented as mean ± SD. Significance was determined by two-tailed unpaired Student’s t test: **p < 0.01; ****p < 0.0001.

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Over Expression, Derivative Assay, Transmission Assay, Electron Microscopy, Plasmid Preparation, Two Tailed Test

Journal: Aging cell

Article Title: Sod2 overexpression preserves myoblast mitochondrial mass and function, but not muscle mass with aging.

doi: 10.1111/j.1474-9726.2009.00477.x

Figure Lengend Snippet: Fig. 5 Measurement of reactive oxygen species (ROS) in myoblasts. (A) Superoxide in mitochondria measured using MitoSox. Left: for reference, wild-type young myoblasts were treated with the increasing concentrations of rotenone (*P < 0.05 compared to wild-type young; **P < 0.05 compared to aged wild-type). (B) Intracellular ROS including hydrogen peroxide (H2O2) and hydroxyl radical (HO·) measured using CM-H2DCFH fluorescence. Left: for reference, intracellular ROS of wild-type young myoblasts treated with increasing concentrations of BSO (*P < 0.05 compared to wild-type young; **P < 0.05 compared to aged wild-type).

Article Snippet: The mitochondrial fraction was extracted from myoblasts grown to confluency on collagen-coated 150-mm dishes using a commercial mitochondria extraction kit (Imgenex, San Diego, CA, USA).

Techniques:

Journal: Chinese Medicine

Article Title: Folium Hibisci Mutabilis extract suppresses M1 macrophage polarization through mitochondrial function enhancement in murine acute gouty arthritis

doi: 10.1186/s13020-025-01081-6

Figure Lengend Snippet: FHME restores mitochondrial stability and improves energy metabolism to reverse LPS/MSU induced M1 polarization. BMDMs were randomly assigned to three groups. Control group was added M-CSF. Model group and FHME group were incubated with M-CSF, LPS/MSU in the presence or absence of 200 µg/mL FHME for 24 h. A Representative transmission electron microscopy images of mitochondrial morphology (× 15,000, × 40,000) in cells. The arrow symbol represented abnormal mitochondria, characterized by bubble change (red arrow), broken ridge and membrane rupture (blue arrow). B Representative Mito-Tracker Red fluorescence images observed the phenotype and morphological changes of mitochondria. C Quantitative analysis of fluorescence intensity of the cells in B. D Alterations in the mitochondrial oxygen consumption rate (OCR) were observed following treatment with FHME. E Statistics analysis of the basal respiration, ATP-linked respiration, maximal respiration, and spare respiration capacity in OCR. F Extracellular acidification rate (ECAR) was measured to quantify glycolytic capacity. G Statistics analysis of the glycolysis, glycolytic capacity, glycolytic reserve, and non-glycolytic acidification in ECAR. Data were calculated by Student’s t-test, *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: The activity of Complex III, also named Coenzyme Q Cytochrome C Reductase, was measured using the Cell Mitochondrial Complex III Activity Assay Kit (#E-BC-K836-M, Elabscience).

Techniques: Control, Incubation, Transmission Assay, Electron Microscopy, Membrane, Fluorescence

Journal: Chinese Medicine

Article Title: Folium Hibisci Mutabilis extract suppresses M1 macrophage polarization through mitochondrial function enhancement in murine acute gouty arthritis

doi: 10.1186/s13020-025-01081-6

Figure Lengend Snippet: FHME promotes UQCRC1 expression to sustain mitochondrial function. A Schematic workflow of the chemical proteomic platform to reveal proteomic changes for FHME treatment in BMDMs. B Venn diagram analysis illustrated the number of differential proteins in each set of experiments, overlapped proteins from triplicated samples were defined as the final list in Model or FHME group. C The Gene Ontology (GO) terms associated with the cellular component were utilized to conduct functional enrichment clustering analysis on the differentially expressed proteins. D A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was conducted on the differential proteins. E The protein–protein interaction (PPI) analysis of mitochondrion pathway-related proteins and oxidative phosphorylation-associated proteins was visualized by Cytoscape. The red region was used to emphasize the higher degree value. F BMDMs were incubated with LPS/MSU with or without FHME for 24 h. Gene levels of UQCRC1, UQCRC2, CYCS and NDUFA4 were determined by qRT-PCR. (G, H) The protein levels of UQCRC1 were detected through western blot ( G ). The quantified protein expression of UQCRC1 in G was normalized to β-actin ( H ). I Representative images depicting the immunohistochemical staining of UQCRC1 in the synovial tissues of mice hind paws. ( J ) Statistics and analysis of the percentage of UQCRC1 positive areas in I. K Cellular mitochondrial complex III activities were assayed by Coenzyme Q-Cytochrome C Reductase Activity Assay Kit after incubating with LPS/MSU with or without FHME for 24 h. Data were calculated by Student’s t-test, *P < 0.05, **P < 0.01, and ***P < 0.001, ns, no significant differences

Article Snippet: The activity of Complex III, also named Coenzyme Q Cytochrome C Reductase, was measured using the Cell Mitochondrial Complex III Activity Assay Kit (#E-BC-K836-M, Elabscience).

Techniques: Expressing, Functional Assay, Phospho-proteomics, Incubation, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Activity Assay