hy 108937 d 64131 mce  (MedChemExpress)

Journal: Cell Death & Disease

Article Title: Inhibition of VDAC1 oligomerization blocks cysteine deprivation-induced ferroptosis via mitochondrial ROS suppression

doi: 10.1038/s41419-024-07216-1

Figure Lengend Snippet: A H1299 cells were transfected with GFP or GFP-VDAC1 for 18 h and subsequently treated with a control or cysteine-deprived medium for the indicated time. B H1299 cells were co-transfected with CTL or VDAC1 siRNAs along with GFP or GFP-VDAC1 for 24 h. C H1299 cells were transfected with GFP or GFP-VDAC1 for 18 h and subsequently treated with 100 μM NSC15364 or 400 μM DIDS for 12 h. A – C The indicated protein levels were estimated by western blot analysis. Band intensities of GFP-VDAC1 were quantified using Image ( J ), normalized to β-actin intensity, and plotted as a fold change relative to the control. D H1299 cells were transfected with GFP or GFP-VDAC1 for 18 h and subsequently treated with a control or cysteine-deprived medium for the indicated time. E H1299 cells were transfected with Flag or Flag-VDAC1 for 24 h. The indicated mRNA levels were estimated by quantitative real-time PCR analysis ( n = 3; *** p < 0.001). F H1299 cells were transfected with Flag or Flag-VDAC1 for 18 h and subsequently treated with control or cysteine-deprived medium for 9 h. G , H H1299 cells were co-transfected with CTL or VDAC1 siRNAs along with Flag or Flag-VDAC1 for 18 h and subsequently treated with control or cysteine-deprived medium for the indicated time ( H ) or 9 h ( G ). I , J H1299 cells were incubated in a control or cysteine-deprived medium with or without NSC15364 or DIDS for the indicated concentration for 24 h. D , H , I , and J Cell viability was measured by MTT assay ( n = 3; *** p < 0.001). K – N H1299 cells were incubated in a control or cysteine-deprived medium with or without 100 μM NSC15364 or 400 μM DIDS for 12 h. F , G A representative flow cytometry of the average MFI ± SD of C11-BODIPY staining ( n = 3; *** p < 0.001, ns not significantly different). K , L A representative flow cytometry histogram of C11-BODIPY staining (upper panels) and the average MFI ± SD of C11-BODIPY staining (low panels) ( n = 3; *** p < 0.001, ns not significantly different). M , N A representative flow cytometry histogram of MitoPeDPP staining (upper panels) and the average MFI ± SD of MitoPeDPP staining (low panels) ( n = 3; *** p < 0.001, ns not significantly different). CTL control, Cys cysteine, MFI mean fluorescence intensity, NSC NSC15364, SD standard deviation.

Article Snippet: DIDS sodium salt (#HY-D0086) and NSC15364 (#HY-108937) were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Transfection, Control, Western Blot, Real-time Polymerase Chain Reaction, Incubation, Concentration Assay, MTT Assay, Flow Cytometry, Staining, Fluorescence, Standard Deviation

Journal: Cell Death & Disease

Article Title: Inhibition of VDAC1 oligomerization blocks cysteine deprivation-induced ferroptosis via mitochondrial ROS suppression

doi: 10.1038/s41419-024-07216-1

Figure Lengend Snippet: A – E H1299 cells were incubated in a control or cysteine-deprived medium with or without 100 μM NSC15364 or 400 μM DIDS for 12 h. A A representative fluorescence microscopy image of MitoSOX red staining. The green fluorescence of MitoBright LT green shows the location of mitochondria (scale bar = 100 μm). B , C A representative flow cytometry histogram of MitoSOX staining (upper panels) and the average MFI ± SD of MitoSOX staining (low panels) ( n = 3; * p < 0.05, *** p < 0.001, ns not significantly different). D , E A representative flow cytometry histogram of TMRE staining. F H1299 cells were incubated in a control or cysteine-deprived medium with or without 100 μM NSC15364 or 400 μM DIDS for 2 h. The oxygen consumption was measured using an extracellular oxygen consumption assay. The dots represent the values of oxygen consumption measured at 1.5-min intervals over a period of 180 min. H1299 cells treated with 75 μM Antimycin A for 2 h were used as a positive control for oxygen consumption rate inhibition, ( n = 3; *** p < 0.001, ns not significantly different). Anti A antimycin A, Cys cysteine, MFI mean fluorescence intensity, NSC NSC15364, SD standard deviation, TMRE tetramethylrhodamine ethyl ester perchlorate.

Article Snippet: DIDS sodium salt (#HY-D0086) and NSC15364 (#HY-108937) were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Incubation, Control, Fluorescence, Microscopy, Staining, Flow Cytometry, Positive Control, Inhibition, Standard Deviation

Journal: Cell Death & Disease

Article Title: Inhibition of VDAC1 oligomerization blocks cysteine deprivation-induced ferroptosis via mitochondrial ROS suppression

doi: 10.1038/s41419-024-07216-1

Figure Lengend Snippet: A H1299 cells were treated with 200 nM RSL3 and 100 μM NSC15364 for 18 h. Cell viability was measured by MTT assay, ( n = 3; * p < 0.05, *** p < 0.001). B – E H1299 cells were treated with 200 nM RSL3 and 100 μM NSC15364 for 9 h. B A representative flow cytometry histogram of C11-BODIPY staining (left panel) and the average MFI ± SD of C11-BODIPY staining (right panel) ( n = 3; *** p < 0.001, ns not significantly different). C A representative flow cytometry histogram of MitoPeDPP staining (left panel) and the average MFI ± SD of MitoPeDPP staining (right panel) ( n = 3; ** p < 0.01, *** p < 0.001). D A representative flow cytometry histogram of MitoSOX staining (left panel) and the average MFI ± SD of MitoSOX staining (right panel) ( n = 3; *** p < 0.001, ns not significantly different). E A representative flow cytometry histogram of TMRE staining (left panel) and the average MFI ± SD of TMRE staining (right panel) ( n = 3; *** p < 0.001, ns not significantly different). F H1299 cells were treated with 200 nM RSL3 and 100 μM NSC15364 for 2 h. The oxygen consumption was measured using an extracellular oxygen consumption assay. The dots represent the values of oxygen consumption measured at 1.5-min intervals over a period of 180 min. H1299 cells treated with 75 μM Antimycin A for 2 h were used as a positive control for oxygen consumption rate inhibition, ( n = 3; *** p < 0.001). Anti A antimycin A, Cys cysteine, MFI mean fluorescence intensity, NSC NSC15364, SD standard deviation, TMRE tetramethylrhodamine ethyl ester perchlorate.

Article Snippet: DIDS sodium salt (#HY-D0086) and NSC15364 (#HY-108937) were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: MTT Assay, Flow Cytometry, Staining, Positive Control, Inhibition, Fluorescence, Standard Deviation

Journal: Cell Death & Disease

Article Title: Inhibition of VDAC1 oligomerization blocks cysteine deprivation-induced ferroptosis via mitochondrial ROS suppression

doi: 10.1038/s41419-024-07216-1

Figure Lengend Snippet: A , B MDA-MB-231 cells were incubated in a control or cysteine-deprived medium with or without NSC15364 or DIDS for the indicated concentration for 24 h. C , D HEYA8 cells were incubated in a control or cysteine-deprived medium with or without NSC15364 or DIDS for the indicated concentration for 24 h. A – D Cell viability was measured by MTT assay ( n = 3). E , G , I , and K MDA-MB-231 cells were incubated in a control or cysteine-deprived medium with or without 100 μM NSC15364 or 400 μM DIDS for 12 h. F , H , J , and L HEYA8 cells were incubated in a control or cysteine-deprived medium with or without 100 μM NSC15364 or 400 μM DIDS for 16 h. E , F A representative flow cytometry histogram of C11-BODIPY staining (upper panels) and the average MFI ± SD of C11-BODIPY staining (low panels) ( n = 3; ** p < 0.01, *** p < 0.001, ns not significantly different). G , H A representative flow cytometry histogram of MitoPeDPP staining (upper panels) and the average MFI ± SD of MitoPeDPP staining (low panels) ( n = 3; * p < 0.05, *** p < 0.001). I , J A representative flow cytometry histogram of MitoSOX. K , L A representative flow cytometry histogram of TMRE. Cys cysteine, MFI mean fluorescence intensity, NSC NSC15364, SD standard deviation, TMRE tetramethylrhodamine ethyl ester perchlorate.

Article Snippet: DIDS sodium salt (#HY-D0086) and NSC15364 (#HY-108937) were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Incubation, Control, Concentration Assay, MTT Assay, Flow Cytometry, Staining, Fluorescence, Standard Deviation