mitochondrial division inhibitor 1 mdivi 1  (MedChemExpress)

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Ameliorates Mitochondria-Dependent Neuronal Apoptosis after Spinal Cord Ischemia-Reperfusion Injury Partially through Inhibiting Oxidative Stress and Promoting Mitophagy

doi: 10.1155/2020/3549704

Figure Lengend Snippet: Mdivi-1, a specific mitophagy inhibitor, partially reversed the protective effect of Sal on spinal cord neurons. (a) Representative images of TUNEL and DAPI costaining of spinal cord neurons that were subjected to control or OGD/R for 12 h. Scale bar, 100 μ m. (b) Quantification of apoptotic (TUNEL+) cells was performed. (c) Representative western blots of cleaved caspase-3/caspase-9, Bcl-2, and Bax expression in neurons that were subjected to control or OGD/R for 12 h. (d) Quantification of activated caspase-3/caspase-9 expression and the ratio of Bcl-2 to Bax normalized to the level of neurons in the control group that were pretreated with DMSO. GAPDH was used as a loading control. (e) Representative images show ROS production in spinal cord neurons and quantification of ROS intensity normalized to the level in control neurons. Scale bar, 100 μ m. Values are expressed as the ratio of fluorescence intensity relative to the control. All values are expressed as mean ± SEM ( n = 4); ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Sal (C14H20O7, CAS#: 10338–51–9, purity > 98%) and mitochondrial division inhibitor 1 (Mdivi-1) (CAS#: 338967-87-6, purity > 98%) were all purchased from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: TUNEL Assay, Western Blot, Expressing, Fluorescence

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Sodium Butyrate Ameliorates Oxidative Stress-Induced Intestinal Epithelium Barrier Injury and Mitochondrial Damage through AMPK-Mitophagy Pathway

doi: 10.1155/2022/3745135

Figure Lengend Snippet: Effects of sodium butyrate (NaB) on oxidative stress, intestinal epithelium barrier, and mitophagy of porcine intestinal epithelial cells (IPEC-J2) after inhibiting mitophagy or AMPK. (a–c) Superoxide dismutase (SOD), glutathione reductase (GSH) activity, and malondialdehyde (MDA) content of IPEC-J2 treated with Mdivi-1 or Compound C (CC). (d) Cellular mitochondrial membrane potential quantification by flow cytometry. (e) Cellular reactive oxygen species (ROS) level quantification by flow cytometry. (f) Intestinal epithelial transepithelial resistance (TER) and fluorescein isothiocyanate dextran 4 kDa (FD4) permeability. (g) Protein expression and quantification of tight junction Claudin-1, Occludin, and ZO-1. ∗ indicates a significant difference compared with the control group ( P < 0.05); # indicates a significant difference compared with the H2O2 group ( P < 0.05); & indicates a significant difference compared with the NaB+H2O2 group ( P < 0.05).

Article Snippet: Trypsin (Beyotime Biotechnology, China), phosphate-buffered saline (PBS) (Bozan Biotechnology, China), penicillin-streptomycin (Solabao Biotechnology, China), fetal bovine serum (Gemini, Australia), CCK-8 kit (Beyotime Biotechnology, China), MitoSpy™ Red CMXRos (Biolegend, USA), immunofluorescence fixative (Sevier Biotechnology, China), Triton-X 100 (Sigma-Aldrich, St. Louis, MO, USA), Glycine (Sinopharm Group, China), DAPI (Beyotime Biotechnology, China), Goat Anti-Mouse IgG Dylight 594 (Earthox, USA), Goat Anti-Mouse IgG Dylight 488 (Earthox, USA), mitochondrial division inhibitor (Mdivi-1) (Selleck, USA), AMPK inhibitor (Compound C, CC) (Selleck, USA), Lipofectamine RNAiMAX, and Lipofectamine 2000 were obtained from Invitrogen (Invitrogen, USA).

Techniques: Activity Assay, Flow Cytometry, Permeability, Expressing

Journal: Conditioning medicine

Article Title: Mitochondrial dynamics and preconditioning in white matter

doi:

Figure Lengend Snippet: (A, B) OGD suppressed Drp-1 levels in MONs in the cytosolic fraction (0.7 ± 0.04%, n = 4) and revealed a corresponding increase in the mitochondrial fraction (2.0 ± 0.4%, n = 6; control 1.0 ± 0.0, n = 7, p< 0.05). Following application of the Drp-1 inhibitor Mdivi-1 (50 μM), cytosolic Drp-1 levels increased to 1.55 ± 0.3%, (n = 3, p < 0.05) and mitochondrial Drp-1 levels dropped to 0.94 ± 0.2% (n = 6, p < 0.05). (C) Confocal microscopy imaging showed that OGD drastically reduced CFP fluorescence in MONs from Thy-1 mito CFP (+) mice. Note that mitochondrial numbers declined and mitochondria became smaller and more round with OGD. Mdivi-1 application before, during, and after OGD preserved CFP pixel intensity and mitochondrial morphology; however, Mdivi-1 preconditioning application failed to preserve CFP fluorescence. (Scale bar, 5μm). n = number of MONs; *p < 0.05, **p < 0.01, one-way ANOVA. Error bars indicate SEM.

Article Snippet: Mitochondrial division inhibitor-1 (Mdivi-1) was purchased from Selleck Chemicals (#S7162, Houston, TX).

Techniques: Confocal Microscopy, Imaging, Fluorescence

Journal: Conditioning medicine

Article Title: Mitochondrial dynamics and preconditioning in white matter

doi:

Figure Lengend Snippet: (A) Mdivi-1 exerts dose-dependent protection of axon function recovery in MONs following OGD. (B) Representative CAP traces at baseline in control ACSF (a), 60 min OGD (b), and recovery (c) conditions are shown for control (green), Mdivi-1 applied as an all-through treatment (pink), and Mdivi-1 applied as a preconditioning treatment (purple). (C) CAP area recovered minimally after OGD (green, 21.6 ± 1.8%, n = 10). Mdivi-1 all-through application (pink) preserved CAP area during OGD and improved CAP area recovery during reperfusion (41 ± 5.0%, n = 7). Mdivi-1 preconditioning treatment (purple) failed to promote axon function recovery (18.6 ± 6.0%, n = 4). Pink and purple horizontal lines represent Mdivi-1 all-through and preconditioning treatment applications, respectively. The inset histograms show quantification of CAP area recovery 2 h after OGD. MONs pretreated with Mdivi-1 (pink) showed improved axon function recovery, whereas the CAP area recovery failed to improve when MONs were preconditioned with Mdivi-1 (purple). n = number of MONs; *p < 0.05, **p < 0.01, one-way ANOVA. Error bars indicate SEM.

Article Snippet: Mitochondrial division inhibitor-1 (Mdivi-1) was purchased from Selleck Chemicals (#S7162, Houston, TX).

Techniques:

Journal: Conditioning medicine

Article Title: Mitochondrial dynamics and preconditioning in white matter

doi:

Figure Lengend Snippet: (A) A representative kymograph created from CFP (+) live mitochondrial imaging shows immotile mitochondria as vertical lines. Note the diagonal lines representing mitochondrial movement in axons in the anterograde direction (blue) as well as the retrograde direction (tan). (B) Quantification of mitochondrial motility from kymographs demonstrated a 50% reduction in mitochondrial motility, both in the anterograde (blue histograms) and retrograde directions (tan histograms), which showed minimal recovery in the anterograde direction reperfusion. Mdivi-1 caused a prominent increase in mitochondrial motility in both directions within 10 min of application (pink histograms), suggesting that inhibition of mitochondrial fission enhanced mitochondrial transport within the myelinated axons. Note that the baseline effect of Mdivi-1 was more prominent in the anterograde direction (pink with blue border histograms). This increase in mitochondrial motility persisted during OGD (60 min) and recovery (20 min) in both directions. n = number of MONs; *p < 0.05, *** p < 0.001, One-way ANOVA with Bonferonni’s post hoc test. (C) Mitochondrial motility analysis showed a similar increase in mitochondrial motility in both directions upon preconditioning application of Mdivi-1 (purple histograms). However, mitochondrial motility was suppressed during OGD and did not improve during the recovery period in either the anterograde or retrograde direction. *** p < 0.001, One-way ANOVA with Bonferonni’s post hoc test.

Article Snippet: Mitochondrial division inhibitor-1 (Mdivi-1) was purchased from Selleck Chemicals (#S7162, Houston, TX).

Techniques: Imaging, Inhibition