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