Journal: Science Advances

Article Title: Plasma iron controls neutrophil production and function

doi: 10.1126/sciadv.abq5384

Figure Lengend Snippet: ( A ) Phenotyping ex vivo of isolated BM neutrophils (obtained from mice treated as in ). Dihydrorhodamine 123 (DHR123) fluorescence as a reporter for cytoplasmic ROS in PMA-stimulated cells. Phagocytosis of GFP-labeled E. coli . FMO, fluorescence minus one. Two-way ANOVA. Mean, quartiles, and range. Bacterial killing of S. aureus , resolved as colony-forming units of S. aureus recovered after coculture with neutrophils. Unpaired t test. Means ± SD. ( B ) Supernatant CCL2 and TNF levels produced by zymosan-stimulated neutrophils measured by enzyme-linked immunosorbent assay (ELISA). Unpaired t test. Mean, quartiles, and range. ( C ) NETosis was evaluated in isolated BM neutrophils after stimulation ex vivo with PMA and ionomycin ± DPI. A minimum of 250 cells from two replicates per sample counted. Two-way ANOVA. Mean, quartiles, and range. Representative microscopy images taken at ×20 magnification. ( D ) NETosis was evaluated in isolated BM neutrophils after stimulation ex vivo with PMA ± apocynin (APO). A minimum of 250 cells from two replicates per sample counted. Two-way ANOVA. Mean, quartiles, and range. Representative microscopy images taken at ×20 magnification. ( E ) SCENITH (single-cell metabolism by profiling translation inhibition) analysis of Ly6G + BM neutrophils from experiment described in , analyzing proportional shifts in O -propargyl-puromycin (OPP) incorporation after ex vivo treatment with metabolic inhibitors as detailed in Materials and Methods. Two-way ANOVA with matching for sample. Reported P value is the effect of mHep treatment. Mean. FAO, fatty acid oxidation; AAO,amino acid oxidation. ( F ) MitoSOX fluorescence as a reporter for mitoROS in ionomycin stimulated isolated BM neutrophils. Two-way ANOVA. Mean, quartiles, and range. ( G ) NETosis was evaluated in isolated BM neutrophils after stimulation ex vivo with ionomycin (iono) ± mitoTEMPO. A minimum of 250 cells from two replicates per sample counted. Two-way ANOVA. Mean, quartiles, and range. Representative microscopy images taken at ×20 magnification.

Article Snippet: A total of 25 μM DPI (Sigma-Aldrich, D2926) was used to block total ROS generation from ionomycin and PMA, 100 μM apocynin (Abcam, ab120615) was used to block ROS generation via NOX2 complex by PMA, and 5 μM mitoTEMPO (Cayman Chemical, 16621) was used to inhibit mitoROS production.

Techniques: Ex Vivo, Isolation, Fluorescence, Labeling, Produced, Enzyme-linked Immunosorbent Assay, Microscopy, Inhibition

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: The role of mitochondrial dysfunction and ROS in mediating hypertension in the RUPP rat model of preeclampsia

doi: 10.1161/HYPERTENSIONAHA.118.11290

Figure Lengend Snippet: Mitochondrial targeted antioxidants attenuate blood pressure in RUPP rats. RUPP rats (n=6) show significantly elevated MAP compared to NP (n=5). Mito Q (500 μM, oral gavage) or MitoTEMPO (1 mg/kg/d, osmotic pumps), improved MAP in RUPPs (n=5-10) compared to untreated RUPPs (n=6) and attenuate any rise in MAP in RUPP rats compared to NP treated with MitoQ (n=6) or MitoTEMPO (n=3). Data are presented as mean ± SEM, *P <0.05, one way ANOVA with Bonferroni post hoc test.

Article Snippet: Moreover, serum from RUPPs treated with MitoQ or MitoTEMPO attenuated mt ROS compared to sera from RUPP control rats ( , 0.2±0.01 or 0.57+0.2, % gated, p<0.05).

Techniques:

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: The role of mitochondrial dysfunction and ROS in mediating hypertension in the RUPP rat model of preeclampsia

doi: 10.1161/HYPERTENSIONAHA.118.11290

Figure Lengend Snippet: Mitochondrial targeted antioxidants improve fetal outcomes in RUPP rats. RUPP rats (n=9-15) show reduced pup weight, placental weight & litter size compared to NP (n=9-10). MitoQ (500 μM, oral gavage) or MitoTEMPO (1 mg/kg/d, osmotic pumps) improved these fetal outcomes in RUPPs (n=4-5) compared to untreated RUPPs (n=9-15) and improve fetal outcomes in RUPP rats compared to NP treated with MitoQ or MitoTEMPO (n=3-6) (A-C). However, MitoQ did not show any improvement in litter size (C). Data are presented as mean ± SEM, *P <0.05, one way ANOVA with Bonferroni posthoc test.

Article Snippet: Moreover, serum from RUPPs treated with MitoQ or MitoTEMPO attenuated mt ROS compared to sera from RUPP control rats ( , 0.2±0.01 or 0.57+0.2, % gated, p<0.05).

Techniques:

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: The role of mitochondrial dysfunction and ROS in mediating hypertension in the RUPP rat model of preeclampsia

doi: 10.1161/HYPERTENSIONAHA.118.11290

Figure Lengend Snippet: ETC activity is reduced in placenta in response to ischemia and may partially be improved with mitochondrial targeted antioxidant treatment. Complex I and Complex IV activities were measured in isolated mitochondrial membranes using spectrophotometer and Oxygraph 2K respectively. Complex I and Complex IV activities were significantly reduced in RUPP placental mitochondria (n=4-5) vs. NP (n=4-6). Mito Q (500 μM, oral gavage) or MitoTEMPO (1 mg/kg/d, osmotic pumps), partially reversed complex I and IV activities in RUPPs (n=3) vs RUPPs (n=4-5) (A&B). Data are presented as mean ± SEM, *P <0.05, t test.

Article Snippet: Moreover, serum from RUPPs treated with MitoQ or MitoTEMPO attenuated mt ROS compared to sera from RUPP control rats ( , 0.2±0.01 or 0.57+0.2, % gated, p<0.05).

Techniques: Activity Assay, Isolation, Spectrophotometry

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: The role of mitochondrial dysfunction and ROS in mediating hypertension in the RUPP rat model of preeclampsia

doi: 10.1161/HYPERTENSIONAHA.118.11290

Figure Lengend Snippet: Mitochondrial targeted antioxidants abrogate mitochondrial ROS production in HUVECs. RUPP (n=7) serum (10%) increases mtROS production in HUVECs vs NP(n=7) serum. Serum (10%) from RUPPs (n=3) treated with MitoQ (500 μM, oral gavage) or MitoTEMPO (1 mg/kg/d, osmotic pumps) significantly reduced mtROS in HUVECs compared to cells that were incubated with RUPP (n=7) serum (A). Histogram showing the mitoSOX staining in HUVECs incubated with NP or RUPP or RUPP+Mito Q or RUPP+MitoTEMPO serum (B). Data are presented as mean ± SEM, *P <0.05, one way ANOVA with Bonferroni post hoc test.

Article Snippet: Moreover, serum from RUPPs treated with MitoQ or MitoTEMPO attenuated mt ROS compared to sera from RUPP control rats ( , 0.2±0.01 or 0.57+0.2, % gated, p<0.05).

Techniques: Incubation, Staining

Journal: Cell

Article Title: TNF Induces Pathogenic Programmed Macrophage Necrosis in Tuberculosis through a Mitochondrial-Lysosomal-Endoplasmic Reticulum Circuit

doi: 10.1016/j.cell.2019.08.004

Figure Lengend Snippet:

Article Snippet: Mitotempo , Cambridge Bioscience , Cat16621-5mg-CAY.

Techniques: Virus, Transformation Assay, Recombinant, Staining, Sequencing, Software, Fluorescence, Imaging

Journal: Cell

Article Title: TDP-43 Triggers Mitochondrial DNA Release via mPTP to Activate cGAS/STING in ALS

doi: 10.1016/j.cell.2020.09.020

Figure Lengend Snippet: TDP-43 Releases mtDNA via the mPTP, Related to Figure 4 (A) Representative western blot of cleaved caspase-3 in TDP-43-overexpressing Mcl1 −/− MEFs 72hrs post doxycycline (Dox) treatment, or treated with ABT-737 to induce apoptosis (t = 4h). (B) Representative western blot of Bak, Bax, TDP-43 and actin from cells in Figure 4 A. (C) iPSC-derived MNs from healthy controls and ALS patients carrying mutations in TDP-43 were stained with Tetramethylrhodamine Methyl Ester (TMRM) to probe mitochondrial membrane potential (mΔΨ) and quantified by FACS analysis (MFI: mean fluorescence intensity). (D) Mitochondrial ROS inhibitors, mitoQ and mitoTEMPO (0.1-1 μM), potently prevent induction of IFNB1 and TNF in TDP-43-ALS iPSC-MNs. (E-F) Representative western blot analysis of cytosolic lysates (0.025% digitonin) and WCL (1x RIPA) from HEK293T cells treated with CsA (12.5 μM) or from (G-H) CRISPR Ppid knockout MEFs transfected with TDP-43-EGFP (WT, A315T and Q331K, 2.5 μg) or untransfected (Ctrl). Cytosolic fraction purity was confirmed using the subcellular markers indicated (I) Treatment with VBIT-4 and CsA (10uM, t = 24h) reduces mtDNA ( MT-ND1 and MT-ND2 ) release into the cytoplasm as performed in Figure S3C, (J) reduces expression of IFNB1 and TNF as determined by qPCR, and (K) reduces production of IFNβ and IP-10 quantified by ELISA. (L) CRISPR Vdac1 knockout MEFs display no TDP-43-induced Ifnb1 and Tnf expression. Data are mean ± SEM from 3-4 independent experiments. P value s were calculated using un-paired t test between healthy control and TDP-43-ALS patient iPSC-MN lines or one-way ANOVA to DMSO or vector control. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: MitoTEMPO (hydrate) , Cayman Chemical , Cat# 16621; CAS 1569257-94-8.

Techniques: Western Blot, Derivative Assay, Staining, Membrane, Fluorescence, CRISPR, Knock-Out, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Cell

Article Title: TDP-43 Triggers Mitochondrial DNA Release via mPTP to Activate cGAS/STING in ALS

doi: 10.1016/j.cell.2020.09.020

Figure Lengend Snippet:

Article Snippet: MitoTEMPO (hydrate) , Cayman Chemical , Cat# 16621; CAS 1569257-94-8.

Techniques: Recombinant, Transfection, Control, Protease Inhibitor, Modification, Knock-Out, Mutagenesis, Activity Assay, Enzyme-linked Immunosorbent Assay, CRISPR, Software

Journal: JCI Insight

Article Title: Aorta- and liver-generated TMAO enhances trained immunity for increased inflammation via ER stress/mitochondrial ROS/glycolysis pathways

doi: 10.1172/jci.insight.158183

Figure Lengend Snippet: ( A ) RNA-Seq data show that TMAO significantly modulated the expression of 4 organelle crosstalk regulators (OCRs) with 2 upregulated genes, PLIN4 and OMA1 , and 2 downregulated genes, JPH1 and PLCH1; TMAO upregulated 2 MitoCarta genes, OGDHL and OMA1 , and downregulated DARS2 . ( B ) Cytoscape analysis showed the connection between TMAO-upregulated mitochondrial gene ( OMA1 ) and 2 ROS regulators (GSEA). ( C ) TMAO induces mitoROS. HAECs were treated with different TMAO concentrations for 2 hours. Then cells were loaded with MitoSOX, and MitoSOX was detected by flow cytometry ( n = 3 for each group); the experiment was repeated 3 times. ( D ) Overproduction of mitoROS contributes to TMAO-induced EC activation. HAECs were treated with TMAO (600 μM) and mitoTempo (1 μM) for 18 hours. ICAM-1 + cell and mean fluorescence intensity (MFI) were detected using flow cytometry analysis ( n = 3 for each group; the experiment was repeated 3 times). Data are represented as the mean ± SEM ( t test; * P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For ICAM-1 measurement, cells were treated with TMAO (600 μM; Sigma Aldrich, 317594), MitoTEMPO (1 μM; Enzon, ALX430-150-M005) , GSK2606414 (1 μM; TOCRIS, 5107), or GSK2656157 (1 μM; MilliporeSigma, 5.04651.0001) ( ) for 18 hours.

Techniques: RNA Sequencing, Expressing, Flow Cytometry, Activation Assay, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Antiproliferation for Breast Cancer Cells by Ethyl Acetate Extract of Nepenthes thorellii x ( ventricosa x maxima )

doi: 10.3390/ijms20133238

Figure Lengend Snippet: MitoSOX and MMP change after EANT treatment. ( A , B ) The concentration effect of EANT on MitoSOX patterns and statistics. Breast cancer cells (MCF7 and SKBR3 were treated with 0 (control with DMSO only), 15, and 25 μg/mL of EANT for 24 h. ( C – F ) Time course effect of EANT on MitoSOX patterns and statistics. Without or with ( C and D ) NAC pretreatment or ( E , F ) MitoTEMPO pretreatment, breast cancer cells (MCF7 and SKBR3) were treated with 25 μg/mL of EANT for 0 (control with DMSO only), 12, and 24 h. Annexin V (+) was counted to apoptosis. ( G , H ) The concentration effect of EANT on MMP patterns and statistics. Breast cancer cells (MCF7 and SKBR3) were treated with EANT for 24 h. ( I , J ) Time course effect of EANT on MMP patterns and statistics. Without or with NAC pretreatment, breast cancer cells (MCF7 and SKBR3) were treated with 25 μg/mL of EANT for 0 (control with DMSO only), 12, and 24 h. For each cell line, treatments labeled without the same lower-case letters indicate significant difference. p < 0.05~0.0001. Data, mean ± SD ( n = 3). (+) or (−) respectively indicates the MitoSOX (+) or MMP (−) population. Positive controls for MitoSOX and MMP were provided in .

Article Snippet: To examine the role of oxidative stress, cells were pretreated with 2 mM ROS inhibitor N -acetylcysteine (NAC) (Sigma-Aldrich; St Louis, MO, USA) [ ] for 1 h. The role of mitochondrial oxidative stress was also examined by pretreating cells with 10 μM of the mitochondrial superoxide inhibitor MitoTEMPO (Cayman Chemical, Ann Arbor, Michigan, USA) [ ] for 1 h. The role of apoptosis was examined by pretreating cells with 20 μM of an apoptosis inhibitor Z-VAD-FMK (Z-VAD) (Selleckchem. com; Houston, TX, USA) [ ] for 2 h.

Techniques: Concentration Assay, Control, Labeling

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: WT (C57BL/6) bone marrow-derived PMNs were infected with S . pneumoniae TIGR4 at various MOIs for 10 minutes and mitochondrial ROS measured using MitoSOX. (A) Gating strategy. (B) The geometric Mean Fluorescent Intensity (MFI) or (C) % of mitochondrial ROS producing cells were determined by flow cytometry. (D) PMNs were treated with vehicle control or the mitochondrial ROS scavenger MitoTEMPO and then mock treated (uninfected) or infected with S . pneumoniae (+Sp) TIGR4 at a MOI of 50. Cells were monitored for intracellular ROS production over the course of 60 minutes using chemiluminescent luminol. (B-D) Representative data shown are from 1 out of 5 separate experiments in which n = 3 technical replicates were used per condition. Line and Bar graphs represent the mean +/-SD. (B and C) * indicates significant differences from uninfected controls and # indicates significant differences between the indicated groups as measured by one-way ANOVA followed by Tukey’s multiple comparison test. (D) * indicates significant differences between infected groups +/- MitoTEMPO as determined by 2-way ANOVA followed by Tukey’s multiple comparisons test.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Derivative Assay, Infection, Flow Cytometry, Control, Comparison

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: (A) WT (C57BL/6) bone-marrow derived PMNs were treated with vehicle control (VC), the mitochondrial ROS scavenger MitoTEMPO or Euk134 or the NADPH oxidase inhibitor DPI and then infected with S . pneumoniae TIGR4. (B) WT (C57BL/6) bone-marrow derived PMNs were treated with vehicle control (VC) or MitoTEMPO and infected with S . pneumoniae D39, 19F, or 23F. The percentage of bacterial killing was determined with respect to no PMN controls under the same treatment conditions. Data are pooled from (A) n = 5 and (B) n = 3 separate experiments. Bar graphs represent the mean +/-SD. * indicates significant differences between the indicated groups as determined by (A) one-way ANOVA followed by Tukey’s multiple comparisons test or (B) unpaired Student’s t-test. n.s. indicates not significant.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Derivative Assay, Control, Infection

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: (A) A2BR -/- (grey) and WT (black) bone-marrow derived PMNs were mock treated (uninfected) or infected with S . pneumoniae (+Sp) TIGR4 at a multiplicity of infection (MOI) of 50 in the presence or absence of MitoTEMPO. Cells were monitored for intracellular ROS production over 60 minutes using luminol. Data are representative from 1 of 3 experiments in which n = 3 technical replicates were used per condition. * indicates significant differences between infected A2BR -/- and WT controls as well as infected A2BR -/- +/- MitoTEMPO treatment as determined by 2-way ANOVA followed by Tukey’s multiple comparisons test. Line graphs represent the mean +/-SD. (B-C) WT (Black) and A2BR -/- (light grey) marrow-derived PMNs were infected with S . pneumoniae TIGR4 at the indicated MOIs for 10 minutes. +MitoTEMPO were treated with the drug prior to infection at an MOI of 50. (B) The % of MitoSOX+ cells as well as (C) the amount of MitoSOX produced (geometric MFI) were determined using flow cytometry. (B-C) Representative data shown are from 1 out of 8 separate experiments in which n = 3 technical replicates were used per condition. Bar graphs represent the mean +/-SD. * indicates significant differences from uninfected controls and # indicates significant differences between the indicated groups as measured by one-way ANOVA followed by Tukey’s multiple comparison test. (D) Fold increases in MitoSOX+ cells upon bacterial infection were calculated by dividing the values of infected conditions by uninfected controls for each mouse strain. Data pooled from eight separate experiments (n = 8 mice/ group) are shown. $ indicates significantly different from 1 as measured by one-sample t-test and * indicates significant differences between the indicated groups as measured by Student’s t-test. (E) WT PMNs were treated with vehicle control (VC) or the A2BR Agonist BAY60-6583 for 30 minutes. Cells were then mock-treated (Uninfected) or infected with S . pneumoniae TIGR4 at a MOI of 10 for 10 minutes. The % of mitochondrial ROS producing cells were determined by flow cytometry. Representative data shown are from 1 out of 5 separate experiments in which n = 3 technical replicates were used per condition. Bar graphs represent the mean +/-SD. * indicates significant differences from uninfected controls and # indicates significant differences between the indicated groups as measured by one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Derivative Assay, Infection, Produced, Flow Cytometry, Comparison, Control

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: WT (C57BL/6) (black) or A2BR -/- (light grey) bone-marrow derived PMNs were treated with vehicle control (VC), the mitochondrial ROS scavenger MitoTEMPO, or the A2BR agonist BAY 60–6583 and then infected with S . pneumoniae TIGR4. The percentage of bacterial killing was determined with respect to no PMN controls under the same treatment conditions. Data are pooled from n = 5 separate experiments. Bar graphs represent the mean +/-SD. * indicates significant differences from VC treated controls for each mouse strain and # indicates significant differences between indicated VC treated WT vs A2BR -/- groups as measured by one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Derivative Assay, Control, Infection, Comparison

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: (A-B) WT C57BL/6 mice treated with vehicle control or the mitochondrial ROS scavenger MitoTEMPO were infected with S . pneumoniae D39 intra-peritoneally. (A) survival as well as (B) bacteremia was followed over time. Data shown are from two separate experiments with n = 5 mice per group. * indicates significant differences as measured by Log-Rank Mantle Cox test (A) and Mann-Whitney test (B). (C-D) A2BR -/- and WT C57BL/6 mice treated with vehicle control or the mitochondrial ROS scavenger MitoTEMPO were infected with S . pneumoniae TIGR4 intra-peritoneally. (C) survival as well as (D) bacteremia and clearance of infection (E) was followed over time. Data shown are from three separate experiments with n = 12 mice per group. * indicates significant differences as measured by Log-Rank Mantle Cox test (C) and Kruskal Wallis test (D). $ indicates significant differences between the mouse groups in the clearance of bacteremia as measured by Fisher’s exact test (E). n.s. indicates not significant.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Control, Infection, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Mitochondrial ROS production by neutrophils is required for host antimicrobial function against Streptococcus pneumoniae and is controlled by A2B adenosine receptor signaling

doi: 10.1371/journal.ppat.1010700

Figure Lengend Snippet: PMNs were isolated from the blood of young healthy donors and (A) infected with S . pneumoniae TIGR4 at the indicated MOIs or mock-treated (uninfected) for 10 minutes. The % of MitoSOX+ cells were determined using flow cytometry. Data shown are from 4 separate donors where each condition was tested in triplicates per donor. Bar graphs represent the mean +/-SD. * indicates significant differences from uninfected controls as determined by one-way ANOVA followed by Dunnett’s multiple comparisons test. (B) PMNs were pre-treated with vehicle control (VC), the general ROS scavenger Ascorbic Acid, the mitochondrial ROS scavenger MitoTEMPO, the NADPH oxidase inhibitor Diphenyleneiodonium chloride (DPI) or the A2B Agonist (BAY 60–6583) and then infected with S . pneumoniae TIGR4. For each donor, the average percent bacterial killing compared to a no PMN control was calculated from triplicate wells per condition. Data from 4 donors are shown. Bar graphs represent the mean +/-SD. * indicates significant differences from VC treated PMNs as determined by one-way ANOVA followed by Dunnett’s multiple comparisons test.

Article Snippet: The effect of Mitochondrial ROS production on host resistance against systemic infection was determined using the mitochondrial ROS scavenger MitoTempo (Cayman Chemical).

Techniques: Isolation, Infection, Flow Cytometry, Control

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: Effect of the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p) on metabolic parameters in rats fed a normal chow (CT) and high fat diet (HFD) fed rats.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: Protein levels of ( A ) fibronectin, ( B ) periostin, ( C ) α-smooth muscle actin (SMA) and ( D ) vimentin in heart from control rats fed a normal chow (CT) and rats fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p). Bars graphs represent the mean ± SEM of 6–8 animals. Protein densitometry was expressed in arbitrary units (AU) once normalized to stain-free gel for protein.* p < 0.05; ** p < 0.01 vs. CT group. † p < 0.05, †† p < 0.01 vs. HFD group.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques: Staining

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: ( A ) Representative microphotographs and ( B ) quantification of total mucin levels in colon from control animals fed a normal chow (CT) and animals fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p) stained with Alcian Blue (AB)/periodic acid-Schiff (PAS) examined by light microscopy (magnification 20×). Bar graphs represent the mean ± SEM of 5–6 animals normalized to for CT group. *** p < 0.001 vs. CT group; †† p < 0.01 vs. HFD group.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques: Staining, Light Microscopy

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: Alpha and beta diversity indexes in rats fed a normal chow (CT) and high fat diet (HFD) and treated with the antioxidant mitochondrial MitoTempo (MT; 0.7 mg/Kg/day i.p).

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: Boxplot showing total relative abundance of reads of the four most abundant taxa at Phylum level in the gut microbiota ( A ) Firmicutes , ( B ) Bacteroidetes , ( C ) Protobacteria and ( D ) Tenericutes in feces from control animals fed a normal chow (CT) and animals fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p). Upper, middle and lower lines represent first quartiles, medians and third quartiles. The whiskers represent a 1.5 * inter-quartile range. Data are expressed as percentage of total reads. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. CT group. † p < 0.05, †† p < 0.01 vs. HFD group.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: LEfSe analysis showing taxonomic differential abundance at family level in feces from control animals fed a normal chow (CT) and animals fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p). ( A ) Significantly different families among CT, HFD and HFD+MT groups. ( B ) Significantly enriched and depleted families between CT and HFD groups. ( C ) Significantly enriched and depleted families between HFD and HFD + MT groups. The length of the horizontal bars represents the LDA score (effect size). p < 0.05; LDA score > 3.0.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: LEfSe analysis showing taxonomic differential abundance at genus level in feces from control animals fed a normal chow (CT) and animals fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p). ( A ) Significantly different genera among CT, HFD and HFD+MT groups. ( B ) Significantly enriched and depleted genera between CT and HFD groups. ( C ) Significantly enriched and depleted genera between HFD and HFD + MT groups. The length of the horizontal bars represents the LDA score (effect size). p < 0.05; LDA score > 3.0.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Antioxidants

Article Title: The Interaction between Mitochondrial Oxidative Stress and Gut Microbiota in the Cardiometabolic Consequences in Diet-Induced Obese Rats

doi: 10.3390/antiox9070640

Figure Lengend Snippet: Boxplot showing total relative abundance of metabolic pathways related to gut microbiota in ( A ) butanoate metabolism, ( B ) propanoate metabolism and ( C ) glutathione metabolism and ( D ) percentage of bacteria involved in lipopolysaccharyde (LPS) production in feces from control animals fed a normal chow (CT) and animals fed a high fat diet (HFD) treated with vehicle or with the mitochondrial antioxidant MitoTempo (MT; 0.7 mg/Kg/day i.p). Upper, middle and lower lines represent first quartiles, medians and third quartiles. The whiskers represent a 1.5 * inter-quartile range. Data are expressed as percentage of total reads. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. CT group. † p < 0.05 vs. HFD group.

Article Snippet: The mitochondrial targeted antioxidant MitoTempo (MT) was obtained from Merck Sigma Aldrich (St. Louis, MO, USA).

Techniques: