Journal: Theranostics

Article Title: Microvascular endothelial metabolic dysfunction drives cerebral edema through bioenergetic failure after ischemia-reperfusion

doi: 10.7150/thno.127083

Figure Lengend Snippet: DDIT4 facilitates the ubiquitination of NDUFS3 resulted in mitochondrial dysfunction. A, Mass spectrometry analysis of proteins pulled down by anti-DDIT4 antibodies identified potential DDIT4-interacting proteins. B, Volcano plot of differentially expressed proteins between mitochondria isolated from cerebral microvascular tissues before and after ischemia-reperfusion. C, Overlapping analysis of differential proteins in mitochondrial proteomic and interacting proteins of DDIT4. D, Heat map showed the expression of 11 overlapping proteins in mitochondrial proteomic. E, Co-immunoprecipitation of NDUFS3 (left) or DDIT4 (right) in ECs (IgG as a control antibody). F, Western blotting of NDUFS3 in indicated cells. Endothelial cell was treated with MG132 6h for harvest. And the quantification of protein was shown in below (n = 6 per group). G, Western blotting of NDUFS3 in cerebral microvascular tissues from conditional DDIT4 knockdown mice. The quantification of protein was shown in below (n = 6 per group). H, Co-IP of NDUFS3 and then western blotted with anti-ubiquitin in DDIT4 overexpression ECs. I, Representative images of staining with mitoSOX (red) and live cell nuclear stain hoechst33342 (blue). J, Phospho-MLKL aggregation in indicated cells. K, IntDen/cell measured using ImageJ (n = 6 per group). L, p-MLKL aggregates/nucleus quantified using Fiji (n = 6 per group). M, The OCR was measured in indicated cells. Data are mean ± SEM.

Article Snippet: The following primary antibody was utilized: Pan Kla (PTM-1401, PTM BIO), H3K9la (PTM-1419, PTM BIO), H3K14la (PTM-1414, PTM BIO), H3K18la (PTM-1406, PTM BIO), H4K5la (PTM-1407, PTM BIO), H4K8la (PTM-1415, PTM BIO), H4K12la (PTM-1411, PTM BIO), H4K16la (PTM-1417, PTM BIO), H3 (PTM-1001, PTM BIO), H4 (PTM-1015, PTM BIO), LDHA (19987-1-AP, Proteintech), β-actin (Abclonal), DDIT4 (10638-1-AP, Proteintech), p-MLKL (ab196436, Abcam), MLKL (37705, Cell Signaling Technology), p-RIP1 (65746S, Cell Signaling Technology), RIP1 (7519-1-AP, Proteintech), p-mTOR (5536S, Cell Signaling Technology), mTOR (66888-1-Ig, Proteintech), NDUFS3 (15066-1-AP, Proteintech), Ubiquitin (43124S, Cell Signaling Technology), ATF4 (60035-1-Ig, Proteintech).

Techniques: Ubiquitin Proteomics, Mass Spectrometry, Isolation, Expressing, Immunoprecipitation, Control, Western Blot, Knockdown, Co-Immunoprecipitation Assay, Over Expression, Staining

Journal: Metabolism Open

Article Title: Supplementation with aspalathin and sulforaphane protects cultured cardiac cells against dyslipidemia-associated oxidative damage

doi: 10.1016/j.metop.2025.100346

Figure Lengend Snippet: The list of TaqMan probes used in the study.

Article Snippet: NADH: Ubiquinone Oxidoreductase Core Subunit S3 , Ndufs3 , Rn01484390_m1.

Techniques: Gene Assay

Journal: Metabolism Open

Article Title: Supplementation with aspalathin and sulforaphane protects cultured cardiac cells against dyslipidemia-associated oxidative damage

doi: 10.1016/j.metop.2025.100346

Figure Lengend Snippet: Aspalathin and sulforaphane increased the mRNA expression of some markers involved in mitochondrial function in cardiomyoblasts exposed to palmitic acid. Briefly, H9c2 cardiomyoblasts were pretreated with 1 μM aspalathin (Asp) and 10 μM sulforaphane (Sul) as well as 2.5 μM Simvastatin (Simva) which was used as a comparative control. Thereafter, cells were co-treated with 0.25 mM palmitic acid (Pal) for an additional 24 h. The mRNA expression levels of peroxisome proliferator-activated receptor gamma coactivator 1-alpha ( Pprgc1α ), nuclear respiratory factor 1 ( Nrf1 ), NADH: ubiquinone oxidoreductase core subunit S3 ( Ndufs3 ), ubiquinol-cytochrome C reductase complex assembly factor 1 ( Uqcc1 ), and uncoupling protein 2 ( Ucp2 ) were quantified ( A, B, C, D, and E , respectively). Results are presented as the mean ± standard error of the mean (SEM) from three independent experiments, each with three technical repeats (n = 3), relative to the experimental control (Ctrl). Comparisons between groups were performed using student’s t -test (and nonparametric test). Statistical significance was represented by ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 compared to the experimental control; and # p < 0.05 compared to the palmitic acid control.

Article Snippet: NADH: Ubiquinone Oxidoreductase Core Subunit S3 , Ndufs3 , Rn01484390_m1.

Techniques: Expressing, Control

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Divergent skeletal muscle mitochondrial phenotype between male and female patients with chronic heart failure

doi: 10.1002/jcsm.12488

Figure Lengend Snippet: Relative mRNA expression levels of several key mitochondrial gene transcripts. Compared with controls, female patients with HFrEF have lower relative mRNA expression for OPA1 (A), PGC‐1α (C), SOD2 (D), NDUFS1 (E), and NDUFS3 (F) ( n = 10 per group for each sex). * P < 0.05 using unpaired Student's t ‐tests. ** P < 0.01 using unpaired Student's t ‐tests. FIS1, mitochondrial fission 1; OPA1, optic atrophy 1; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; NDUFS3, NADH:ubiquinone oxidoreductase core subunit S3; PGC‐1α, peroxisome proliferator‐activated receptor γ coactivator‐1α; SOD2, superoxide dismutase 2.

Article Snippet: Primers were purchased from Qiagen including PGC‐1α, superoxide dismutase 2 (SOD2), FIS1, OPA1, NADH:ubiquinone oxidoreductase core subunit S1 (NDUFS1), and NADH:ubiquinone oxidoreductase core subunit S3 (NDUFS3).

Techniques: Expressing

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Divergent skeletal muscle mitochondrial phenotype between male and female patients with chronic heart failure

doi: 10.1002/jcsm.12488

Figure Lengend Snippet: Correlations between gene transcript expression levels with peak pulmonary oxygen uptake (V̇O 2peak ) and measures of mitochondrial function and content in heart failure with reduced ejection fraction patients

Article Snippet: Primers were purchased from Qiagen including PGC‐1α, superoxide dismutase 2 (SOD2), FIS1, OPA1, NADH:ubiquinone oxidoreductase core subunit S1 (NDUFS1), and NADH:ubiquinone oxidoreductase core subunit S3 (NDUFS3).

Techniques: Expressing