vdac1 inhibitor vbit12  (Selleck Chemicals)

Journal: Cell Death Discovery

Article Title: Rafoxanide disrupts mitochondrial homeostasis through VDAC1 modulation in colorectal cancer cells

doi: 10.1038/s41420-026-02986-3

Figure Lengend Snippet: Representative western blots showing VDAC1 monomeric and dimeric forms, together with β-actin, in HCT116 cells pre-treated or not with the specific VDAC1 inhibitor VBIT12 (40 µM) for 2 h and then stimulated with rafoxanide (RFX, 2.5 μM) or DMSO (vehicle) for 5, 15, 30, and 60 min. One of 3 independent experiments in which similar results were obtained is shown.

Article Snippet: HCT116 cells were pre-treated for 2 h with the VDAC1 inhibitor VBIT12 (40 μM, #S8936, Selleck Chemicals LLC, Houston, Texas) to block VDAC1 oligomerization or for 1 h with N-acetylcysteine (NAC, 1 mM, #A0150000, Sigma Aldrich) to decrease reactive oxygen species (ROS) level.

Techniques: Western Blot

Journal: Cell Death Discovery

Article Title: Rafoxanide disrupts mitochondrial homeostasis through VDAC1 modulation in colorectal cancer cells

doi: 10.1038/s41420-026-02986-3

Figure Lengend Snippet: A Detection of total cellular reactive oxygen species (ROS) by fluorescence intensity measurement in HCT116 cells pre-treated or not with the ROS scavenger NAC (1 mM) for 1 h and then incubated with fluorescence probe DCFDA. After 30 min, cells were washed and stimulated with rafoxanide (RFX, 2.5 μM) or DMSO (vehicle). Fluorescence intensity was measured at the baseline and after 5, 15, 30, and 60 min upon stimulation. Differences among groups were compared using one-way analysis of variance (ANOVA) followed by the Tukey’s post hoc test (* P ≤ 0.05, ** P ≤ 0.01). One of 3 independent experiments where similar results were obtained. B Representative western blots showing VDAC1 monomeric and dimeric forms, together with β-actin, in HCT116 cells pre-treated or not with the ROS scavenger NAC (1 mM) for 1 h and then stimulated with rafoxanide (RFX, 2.5 μM) or DMSO (vehicle) for 1 h. One of 3 independent experiments in which similar results were obtained is shown. C Left panel : representative flow cytometry dot plots showing JC-1 fluorescence in HCT116 cells treated as indicated in panel ( B ). JC-1 aggregates indicate polarized mitochondria, whereas JC-1 monomers indicate mitochondrial membrane depolarization. Numbers indicate the percentage of cells in the designated quadrants. Right panel : quantification of the fraction of cells with depolarized mitochondria. Values are mean ± SEM of 3 independent experiments. Differences among groups were compared using one-way ANOVA followed by Tukey’s post hoc test (** P ≤ 0.01, *** P ≤ 0.001). D , E Complex I ( D ) and complex III ( E ) activity assays were performed spectrophotometrically in HCT116 cells stimulated with rafoxanide (RFX, 2.5 μM) or DMSO (vehicle) for 5 and 15 min. Values are mean ± SEM of 3 independent experiments. Differences among groups were compared using one-way ANOVA followed by Tukey’s post hoc test (* P ≤ 0.05, ** P ≤ 0.01). F Calcium mobilization was measured by flow cytometry in HCT116 cells stained with the 520-AM dye for 30 min, washed, and stimulated with rafoxanide (RFX, 2.5 μM), DMSO (vehicle), or ionomycin (positive control). Samples were acquired by flow cytometry at the baseline and immediately after stimulation. One of 3 independent experiments in which similar results were obtained is shown.

Article Snippet: HCT116 cells were pre-treated for 2 h with the VDAC1 inhibitor VBIT12 (40 μM, #S8936, Selleck Chemicals LLC, Houston, Texas) to block VDAC1 oligomerization or for 1 h with N-acetylcysteine (NAC, 1 mM, #A0150000, Sigma Aldrich) to decrease reactive oxygen species (ROS) level.

Techniques: Fluorescence, Incubation, Western Blot, Flow Cytometry, Membrane, Activity Assay, Staining, Positive Control

Journal: bioRxiv

Article Title: STING and VDAC inhibitors attenuate inflammation and ineffective erythropoiesis caused by an altered metabolome in the Nan (EKLF/E339D) mouse model of neonatal anemia

doi: 10.64898/2025.12.11.693792

Figure Lengend Snippet: VDAC1 inhibitor treatments significantly improves anemic hallmarks in adult Nan/+ mice. (A) Schematic diagram showing VDAC inhibitor (DMSO or VBIT4 or VBIT 12, 5 mg/kg) treatment. Oral gavage was used to introduce mice with DMSO (control) or VBIT4 or 12, and blood samples analyzed at various weeks as indicated (left top), followed by total harvest of bone marrow (BM) at the end for blood parameter. Quantification of RBC numbers (top middle), hematocrit (left bottom) and reticulocyte counts (right bottom). (B) Representative flow plot of BM cells isolated from DMSO or VBIT 4-treated mice and were analyzed for Ter119+ (left) and quantification (right). (C) Representative flow plot of bone marrow cells isolated from DMSO or VBIT 4-treated mice and were analyzed for red cell maturation and gated for G1-G5 as indicated (bottom), and quantification (bottom). (D) mitoROS analyzed in BM Ter119 + cells treated with DMSO or VBIT 4; histograms (left) and quantification (right) represented by Geo.mean. (E) MMP analyzed in BM Ter119 + cells treated with DMSO or VBIT 4; histograms (left) and quantification (right) represented by Geo.mean. (F) Serum IFN-α and -ß levels measured using ELISA in mice treated with DMSO or VBIT4. (G) Spleens were isolated from DMSO or VBIT4- or VBIT12-treated mice and weighed. Size is shown ( left ) and weight is quantified ( right ). Each experiment is an average (n=5). In all panels, data are presented as mean ± S.E.M. ( ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, and ****p < 0.0001). (H) Model : Nan/+ fetal livers (E13.5) show ectopic and hypomorphic gene expression patterns due to the presence of the E339D variant. These cells are aberrantly enriched in L-carnitine and other metabolites. Their mitochondrial morphology is hyperfused and exhibit enhanced, but variable, degrees of disorganization, such as loss of cristae and detachment of inner membrane from outer membrane. Oligomerized VDAC1 mediates mtDNA release to the cytosol, which activates the mtDNA-cGAS-STING signaling pathway leading to increased transcripts for IFNs, interferon response genes (ISGs), and several inflammatory chemokine and cytokines. This mitochondrial dysregulation induces an inflammatory environment that inhibits proper erythroid maturation and causes ineffective erythropoiesis in Nan/+ embryos which persists into adulthood. Inhibition of the mtDNA-cGAS-STING module such as VDAC and STING with small molecules lowers the inflammatory response specifically in the Nan/+ erythroid cells and restores ineffective erythropoiesis seen in embryos fetal liver cells and in adults. (Model drawn via Biorender).

Article Snippet: VBIT-4 and VBIT12 was obtained from Selleckchem.

Techniques: Introduce, Control, Isolation, Enzyme-linked Immunosorbent Assay, Gene Expression, Variant Assay, Membrane, Inhibition