mitochondrial respiratory chain complex i v activity assay kit  (Beijing Solarbio Science)

Journal: International Journal of Oncology

Article Title: SLC25A1 promotes lymph node metastasis of esophageal squamous cell carcinoma by regulating lipid metabolism

doi: 10.3892/ijo.2025.5721

Figure Lengend Snippet: SLC5A1 promotes lipid synthesis and affects oxidative phosphorylation of ESCC cells. (A) Intracellular lipids were stained with fluorescent lipophilic dye BODIPY 493/503. Levels of (B) free fatty acids, (C) triglycerides, and (D) cholesterol. Following glucose starvation, (E) oxygen consumption rate and (F) activity of mitochondrial respiratory chain complexes in ESCC cells was determined. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. SLC5A1, solute carrier family 25 member 1; ESCC, esophageal squamous cell carcinoma; ns, not significant; NC, Negative control; sh, short hairpin.

Article Snippet: Mitochondrial respiratory chain activity was evaluated via the Mitochondrial Respiratory Chain Complex I-V Activity Assay kit (BC0515, BC3230, BC3240, BC0945, BC1445) (Beijing Solarbio Science & Technology Co., Ltd.), according to the manufacturer's instructions.

Techniques: Phospho-proteomics, Staining, Activity Assay, Negative Control

Journal: Cell & Bioscience

Article Title: Ubiquitinated AIF is a major mediator of hypoxia-induced mitochondrial dysfunction and pulmonary artery smooth muscle cell proliferation

doi: 10.1186/s13578-022-00744-3

Figure Lengend Snippet: Hypoxia results in decreased AIF expression. A Activity of mitochondrial respiratory chain complexes I–V after exposure to hypoxia, with complex I showing the most severe damage (n = 6). B Schematic structural model of AIF protein. C Decreased AIF expression was found in plasma from the hypoxic group (n = 5). D , E AIF protein and RNA levels in the lung tissues of the hypoxic model (n = 8). F , G The location of AIF in the smooth muscle layer of lung tissues from the hypoxic model was determined by immunofluorescence ( F , Scale bar = 100 μm) and immunohistochemical staining analysis ( G , Scale bar = 200 μm) (n = 3). H , I Time course of AIF protein and RNA levels in PASMCs 0, 24, 48, and 72 h after hypoxia treatment. J Subcellular distribution of AIF in PASMCs as determined by immunofluorescence analysis. Scale bars: 50 μm (n = 3). All data are presented as the means ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001; Nor normoxia, Hyp hypoxia

Article Snippet: The activities of mitochondrial respiratory chain complex I–V were detected according to the instructions of Micro Mitochondrial Respiratory Chain Complex I–V Activity Assay Kit respectively (Solarbio Life Sciences, Beijing, China).

Techniques: Expressing, Activity Assay, Clinical Proteomics, Immunofluorescence, Immunohistochemical staining, Staining, Standard Deviation

Journal: Cell & Bioscience

Article Title: Ubiquitinated AIF is a major mediator of hypoxia-induced mitochondrial dysfunction and pulmonary artery smooth muscle cell proliferation

doi: 10.1186/s13578-022-00744-3

Figure Lengend Snippet: Hypoxic mitochondrial dysfunction in PASMCs is mediated by AIF. A Activity of complexes I, II and V after pretreatment with AIF overexpressed plasmid (n = 5–6). B Representative images and averaged values of mitochondrial ROS (n = 6). Scale bars: 50 μm. C The oxygen consumption rates of cultured rat PASMCs, maximal respiration, spare respiratory capacity and derived mitochondrial ATP production were calculated following oligomycin and FCCP treatment (n = 3). D Glycolysis and glycolytic reserve following treatment with 10 mM glucose and 1 μM oligomycin were measured in cultured PASMCs, and nonglycolytic acidification after treatment with 100 mM 2-deoxyglucose was detected (n = 3). E Western blot analysis of HK II, PKM2 and PDH protein expression in PASMCs in the presence or absence of hypoxia (n = 6–8). All data are presented as the means ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001; Nor normoxia, Hyp hypoxia, NC negative control

Article Snippet: The activities of mitochondrial respiratory chain complex I–V were detected according to the instructions of Micro Mitochondrial Respiratory Chain Complex I–V Activity Assay Kit respectively (Solarbio Life Sciences, Beijing, China).

Techniques: Activity Assay, Plasmid Preparation, Cell Culture, Derivative Assay, Western Blot, Expressing, Standard Deviation, Negative Control