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