mitochondrial complex activity detection kit  (Elabscience Biotechnology)

Journal: Redox Biology

Article Title: Oxidative modification of G-quadruplex triggers CLIC4-associated mitochondrial dysfunction to promote glioblastoma progression

doi: 10.1016/j.redox.2025.103917

Figure Lengend Snippet: CLIC4 maintains redox homeostasis of glioblastoma cells via regulating mitochondrial ROS A. Subcellular localization of CLIC4 protein assessed by western blotting in U87MG and A172 cells. B and C. Cellular colocalization of CLIC4 with mitochondria in U87MG and A172 as demonstrated by immunofluorescence using CLIC4 and COXIV antibody. D and E. Representative confocal images of U87MG and A172 cells after silencing CLIC4 stained with MitoSOX. F and G. Intracellular ROS levels in U87MG and A172 cells with CLIC4 knockdown were detected by Flow cytometry analysis combined with DCFH-DA staining. H. CLIC4 knockdown with shCLIC4_1# and shCLIC4_2# increases mitochondrial membrane potential in U87MG and A172 cell by flow cytometry. I. Representative images of confocal microscopy combined with immunochemical staining showed γ-H2AX foci formed in the nuclei of the U87MG and A172 cells with CLIC4 knockdown. J. Representative images of confocal microscopy combined with immunochemical staining showed that 8-OHdG level in U87MG and A172 cells with CLIC4 knockdown. K. CLIC4 knockdown with shCLIC4_1# and shCLIC4_2# decreases the number of mitochondria in U87MG and A172 cells. L. Immunoblot of mitochondrial marker proteins including VDAC1, COX IV, MTCO2. M. Mitochondrial complex activity in the U87MG and A172 cells after silencing CLIC4 were assessed via colorimetric method.

Article Snippet: The mitochondrial complex activity detection kit (colorimetric method) from Elabscience (Wuhan, China) was used to measure mitochondrial complex activity, and was performed according to the manufacturer's instructions.

Techniques: Western Blot, Immunofluorescence, Staining, Knockdown, Flow Cytometry, Membrane, Confocal Microscopy, Marker, Activity Assay

Journal: Redox Biology

Article Title: Oxidative modification of G-quadruplex triggers CLIC4-associated mitochondrial dysfunction to promote glioblastoma progression

doi: 10.1016/j.redox.2025.103917

Figure Lengend Snippet: CLIC4 knockdown inhibits GBM growth in an orthotopic mouse model. A. Luciferase image of tumors from respective experimental groups. B. Quantitative analysis was performed to compare the tumor size of shNC and shCLIC4_2# groups by luciferase image at 24 days (n = 6). C. H&E staining for mice brain from shNC and shCLIC4_2# groups,and tumor size (highlighted in red) was quantified for statistical analysis (n = 6). D-G. Images (D) and statistical results (E, F, G) of immunohistochemical staining score for CLIC4, Ki67, 8-OHdG in tumors from each group (n = 6). H and I. The protein expression levels of CLIC4 and COX IV in mouse tumor tissues were detected via immunoblotting (n = 4). J. Schematic model showing the mechanism by which oxidative DNA modification triggers CLIC4-associated mitochondrial dysfunction to promote glioblastoma progression.

Article Snippet: The mitochondrial complex activity detection kit (colorimetric method) from Elabscience (Wuhan, China) was used to measure mitochondrial complex activity, and was performed according to the manufacturer's instructions.

Techniques: Knockdown, Luciferase, Staining, Immunohistochemical staining, Expressing, Western Blot, Modification

Journal: Advanced Functional Materials

Article Title: Biosponge‐Armored Nanodots Restore Redox‐Calcium Homeostasis to Mitigate Reperfusion‐Induced Injury in Ischemic Stroke

doi: 10.1002/adfm.202503183

Figure Lengend Snippet: Figure 5. Protective effects of HIr-PS on mitochondrial dysfunction and endoplasmic reticulum stress. a) Schematic diagram illustrates the dynamic relationship between alterations in the ETC and the interplay between calcium ions and ROS generation (Created with BioRen der.com). b) Representative images depict the mPTP status in SH-SY5Y cells following 6 h of coincubation with H2O2 and various treatments, assessed using the mPTP fluorescence assay lit. c) Quantification of mPTP fluorescence intensity (n = 3). d) Assessment of respiratory chain Complex I (n = 4) and e) Complex III (n = 3) activities in H2O2-stimulated SH-SY5Y cells after different treatments. f) NAD+, g) NADH, and h) ATP levels in H2O2-stimulated SH-SY5Y cells treated with different interventions (n = 3). i) Detection of Cyt c release in SH-SY5Y cells treated with different concentrations of HIr-PS. Green fluorescence indicates Cyt c by immunostaining with an anti-Cyt c antibody, whereas red fluorescence shows the MitoTracker deep red dye in the stained mitochondria. j) Colocalization of Cyt c and mitochondria within individual SH-SY5Y cells, with the white line in Figure 5i indicating the trajectory of its distribution. k) Schematic illustration of the mechanism by which HIr-PS treatment alleviates ERS and restores mitochondrial function, effectively disrupting the ROS and calcium feedback loop (Created with BioRen der.com). l) qPCR results for ERS-related genes and fibrosis marker genes in SH-SY5Y cells before and after HIr-PS treatment (n = 3). All data are presented as mean ± SD. *p < 0.05 **p < 0.01, ***p < 0.001 (one-way ANOVA with Tukey’s post hoc test).

Article Snippet: The activities of respiratory chain Complexes I and III were measured using a mitochondrial respiratory chain complex activity detection kit (Solarbio, Beijing, China), following the manufacturer’s instructions.

Techniques: Immunostaining, Staining, Marker