Journal: Frontiers in Pharmacology

Article Title: Changes in PGC‐1α/SIRT1 Signaling Impact on Mitochondrial Homeostasis in Amyloid-Beta Peptide Toxicity Model

doi: 10.3389/fphar.2020.00709

Figure Lengend Snippet: Acute effects of AβOs on total levels of proteins that regulate mitochondrial dynamics. (A) Mfn1 and (B) DRP1 western blot of lysates from PC12 cells treated with FCCP (10 μM, 2 h) and AβOs (0.5 μM, 1 and 2 h). A dotted line on the western blot indicates different regions of the same gel. (C, D) Quantification of Mfn1 and DRP1 levels normalized to beta actin. Data are represented as mean ± SEM. *p < 0.05, **p < 0.01 compared between the control group. One-way ANOVA with the Dunnett's multiple comparisons test was used for all statistical analyses. Mfn1, mitofusin 1; DRP1, dynamin-related protein 1. (n = 4–5 for each group) (original gel blot are provided on ).

Article Snippet: Samples were incubated for 1 h at RT with the following primary antibodies: SIRT1 (mouse 1:300, Novus Biologicals, IF3), PGC-1α (rabbit 1:400, Novus Biologicals, NBP1-04676), Ser-46 SIRT1 (Sigma 1:200, SAB4301426), DRP1 (rabbit 1:200, Novus Biologicals, NB 110-55288), and Mfn1 (rabbit 1:200, Novus Biologicals, NBP1-51841).

Techniques: Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Changes in PGC‐1α/SIRT1 Signaling Impact on Mitochondrial Homeostasis in Amyloid-Beta Peptide Toxicity Model

doi: 10.3389/fphar.2020.00709

Figure Lengend Snippet: Acute effects of AβOs on immunoreactivity of proteins that regulate mitochondrial dynamics. Representative epifluorescence images of (A) Mfn1 and (B) DRP1 immunoreactivity in PC-12 cells control and treated with AβOs (0.5 μM) for 1 and 2 h. Quantification of (C) Mfn1 and (D) DRP1 immunoreactivity (intensity), under the same experimental conditions. Scale bars: 20 μm. Data are represented as mean ± SEM. *p < 0.05, ***p < 0.001 compared between the control group. One-way ANOVA with the Dunn's multiple comparisons test was used for all statistical analyses. Mfn1, mitofusin 1; DRP1, dynamin-related protein 1. (n=3–6 for each group, N= 56–103) (entire inmunocytochemistry for Mfn1 and DRP1 with control are provided on ).

Article Snippet: Samples were incubated for 1 h at RT with the following primary antibodies: SIRT1 (mouse 1:300, Novus Biologicals, IF3), PGC-1α (rabbit 1:400, Novus Biologicals, NBP1-04676), Ser-46 SIRT1 (Sigma 1:200, SAB4301426), DRP1 (rabbit 1:200, Novus Biologicals, NB 110-55288), and Mfn1 (rabbit 1:200, Novus Biologicals, NBP1-51841).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Changes in PGC‐1α/SIRT1 Signaling Impact on Mitochondrial Homeostasis in Amyloid-Beta Peptide Toxicity Model

doi: 10.3389/fphar.2020.00709

Figure Lengend Snippet: Model of alterations induced on SIRT1/PGC-1α pathway by AβOs. (A) . Healthy neuronal conditions were energy depletion and/or decreased catabolic rates can be sensed by SIRT1 promoting the PGC-1α deacetylation (1). Transcription and co-activates of factors like NRF- 1/2 for the expression of nuclear-encoded mitochondrial genes and dynamic mitochondrial proteins, requires of PGC-1α translocation (2). Coordinated mitochondrial dynamics (3, fission/fusion), depends of adequate expression of Mfn1, Drp1. (B) . In AβOs treated neurons, the PGC-1α is unable to be deacetylated and to translocate to the nucleus (4). The expression of key genes is loss (5), and imbalance between fusion and fission to promote the granular mitochondrial phenotype (6) and neuronal death.

Article Snippet: Samples were incubated for 1 h at RT with the following primary antibodies: SIRT1 (mouse 1:300, Novus Biologicals, IF3), PGC-1α (rabbit 1:400, Novus Biologicals, NBP1-04676), Ser-46 SIRT1 (Sigma 1:200, SAB4301426), DRP1 (rabbit 1:200, Novus Biologicals, NB 110-55288), and Mfn1 (rabbit 1:200, Novus Biologicals, NBP1-51841).

Techniques: Expressing, Translocation Assay

Journal: Journal of Cardiovascular Pharmacology

Article Title: Hydrogen Sulfide Diminishes Activation of Adventitial Fibroblasts Through the Inhibition of Mitochondrial Fission

doi: 10.1097/fjc.0000000000001250

Figure Lengend Snippet: FIGURE 2. Drp1-mediated mitochondrial fission is required for TGF-b1–induced AF activation. AFs were transfected with Drp1 siRNA or scrambled siRNA, followed by treatment with TGF-b1 (10 ng/mL) for 24 hours A, B, The expression of Drp1 was determined by western blotting. C, Micrographs of mitochondrial morphology stained by MitoTracker Red of AFs. Scale bar =10 mm. D, Quantification of mitochondrial morphology. E, Quantitative analysis for collagen I. F, The expression of collagen I and a-SMA was determined by western blotting. G, Quantitative analysis for a-SMA. H, I, Quantification by cell counting Kit-8 assay and cell counting. J, Representative images of transwell migration assay. Scale bar =100 mm. K, Quantification of migrated cells. Values are represented as means 6 standard error of the mean (n = 3); *P , 0.05 versus control group; #P , 0.05 versus TGF-b1 group.

Article Snippet: Published by Wolters Kluwer Health, Inc. the enhanced chemiluminescence western blotting detection system (Amersham Pharmacia, Deisenhofen, Germany). siRNA Transfection Specific small interfering RNA (siRNA) targeting ROCK1 and Drp1 was purchased from Santa Cruz Biotechnology.

Techniques: Activation Assay, Transfection, Expressing, Western Blot, Staining, Cell Counting, Transwell Migration Assay, Control

Journal: Journal of Cardiovascular Pharmacology

Article Title: Hydrogen Sulfide Diminishes Activation of Adventitial Fibroblasts Through the Inhibition of Mitochondrial Fission

doi: 10.1097/fjc.0000000000001250

Figure Lengend Snippet: FIGURE 3. ROCK1 is involved in mediating TGF-b1–induced mitochondrial fission. AFs were transfected with ROCK1 siRNA or scrambled siRNA, followed by treatment with TGF-b1 (10 ng/mL) for 24 hours A, B, The expression of ROCK1 was determined by western blotting. C, Micrographs of mitochondrial morphology stained by MitoTracker Red of AFs. Scale bar =10 mm. D, Quantification of mitochondrial morphology. E, The expression of pDrp1 and Drp1 were determined by western blotting. F, Quantitative analysis for pDrp1. G, The expression of collagen I and a-SMA was determined by western blotting. H, Quantitative analysis for collagen I. I, Quantitative analysis for a-SMA. J, K, Quantification by cell counting Kit-8 assay and cell counting. L, Representative images of transwell migration assay. Scale bar =100 mm. M, Quantification of migrated cells. Values are repre- sented as means 6 standard error of the mean (n = 3); *P , 0.05 versus control group; #P , 0.05 versus TGF-b1 group.

Article Snippet: Published by Wolters Kluwer Health, Inc. the enhanced chemiluminescence western blotting detection system (Amersham Pharmacia, Deisenhofen, Germany). siRNA Transfection Specific small interfering RNA (siRNA) targeting ROCK1 and Drp1 was purchased from Santa Cruz Biotechnology.

Techniques: Transfection, Expressing, Western Blot, Staining, Cell Counting, Transwell Migration Assay, Control

Journal: Antioxidants

Article Title: Pimozide and Imipramine Blue Exploit Mitochondrial Vulnerabilities and Reactive Oxygen Species to Cooperatively Target High Risk Acute Myeloid Leukemia

doi: 10.3390/antiox10060956

Figure Lengend Snippet: IB suppresses phosphorylation of DRP1 in Flt3-ITD + cells. ( A ) A panel of genes related to the ER–mitochondria interface were assayed by real-time qRT-PCR. The panel included Fis1, Ip3r, Mfn1, Drp1, Grp75, Mtorc2, Vdac1, and Bap31 ( n = 3 for all except Drp1 where n = 7). p values were calculated relative to OCI-AML3 cells. ( B ) Western blot analysis for total Drp1 protein with β-actin as the loading control. ( C ) Western blot analysis for phospho-Drp1 protein with β-actin as the loading control. ( D , E ) Densitometry analysis was performed and quantitation is shown for replicates of the Western blots in panels C and D. n = 3 and p values were calculated with comparisons as shown between either OCI-AML3 cells or with or without treatment. * p < 0.05; ** p < 0.01.

Article Snippet: Antibodies for Western blot were obtained from the following: Anti-STAT5 (Phospho-STAT5 Y694) antibody (Abcam, Cambridge, UK), Phospho-Akt (Ser473) (D9E), Phospho-Akt (Thr308) (D25E6) (Cell Signaling), Phospho-DRP1 (Ser637) (D3A4) (Cell Signaling, Danvers, MA, USA), DRP1 Antibody (Novus Biologicals), p38alpha MAPK antibody and Phospho-p38alpha MAPK (Cell Signaling, Danvers, MA, USA).

Techniques: Phospho-proteomics, Quantitative RT-PCR, Western Blot, Control, Quantitation Assay

Journal: Journal of Alzheimer's disease : JAD

Article Title: Mitochondria-Division Inhibitor 1 Protects Against Amyloid-β induced Mitochondrial Fragmentation and Synaptic Damage in Alzheimer’s Disease

doi: 10.3233/JAD-170051

Figure Lengend Snippet: Summary of quantitative real-time RT-PCR oligonucleotide primers used in measuring mRNA expression in mitochondrial dynamics, mitochondrial biogenesis, and synaptic genes in in N2a cells treated with Aβ42, Mdivi1+Aβ42, Aβ42+Mdivi1 relative to untreated N2a cells

Article Snippet: An ID fine-band command was used to locate and to scan the bands in each lane and to record the readings. table ft1 table-wrap mode="anchored" t5 caption a7 Marker Primary Antibody – Species and Dilution Purchased from Company, City & State Secondary Antibody, Dilution Purchased from Company, City & State Drp1 Rabbit Polyclonal 1:500 Novus Biological, Littleton, CO Donkey Anti-rabbit HRP 1:10,000 GE Healthcare Amersham, Piscataway, NJ Fis1 Rabbit Polyclonal 1:500 MBL International Corporation-life.

Techniques: Quantitative RT-PCR, Expressing, Sequencing

Journal: Journal of Alzheimer's disease : JAD

Article Title: Mitochondria-Division Inhibitor 1 Protects Against Amyloid-β induced Mitochondrial Fragmentation and Synaptic Damage in Alzheimer’s Disease

doi: 10.3233/JAD-170051

Figure Lengend Snippet: Summary of antibody dilutions and conditions used in the immunoblotting analysis of mitochondrial dynamics, mitochondrial biogenesis, and synaptic proteins in N2a cells treated with Aβ42, Mdivi1+Aβ42, Aβ42+Mdivi1 relative to untreated N2a cells

Article Snippet: An ID fine-band command was used to locate and to scan the bands in each lane and to record the readings. table ft1 table-wrap mode="anchored" t5 caption a7 Marker Primary Antibody – Species and Dilution Purchased from Company, City & State Secondary Antibody, Dilution Purchased from Company, City & State Drp1 Rabbit Polyclonal 1:500 Novus Biological, Littleton, CO Donkey Anti-rabbit HRP 1:10,000 GE Healthcare Amersham, Piscataway, NJ Fis1 Rabbit Polyclonal 1:500 MBL International Corporation-life.

Techniques: Western Blot

Journal: Journal of Alzheimer's disease : JAD

Article Title: Mitochondria-Division Inhibitor 1 Protects Against Amyloid-β induced Mitochondrial Fragmentation and Synaptic Damage in Alzheimer’s Disease

doi: 10.3233/JAD-170051

Figure Lengend Snippet: mRNA fold changes in N2a cells treated with Aβ42 and Mdivi1

Article Snippet: An ID fine-band command was used to locate and to scan the bands in each lane and to record the readings. table ft1 table-wrap mode="anchored" t5 caption a7 Marker Primary Antibody – Species and Dilution Purchased from Company, City & State Secondary Antibody, Dilution Purchased from Company, City & State Drp1 Rabbit Polyclonal 1:500 Novus Biological, Littleton, CO Donkey Anti-rabbit HRP 1:10,000 GE Healthcare Amersham, Piscataway, NJ Fis1 Rabbit Polyclonal 1:500 MBL International Corporation-life.

Techniques:

Journal: Molecular and Cellular Biology

Article Title: Retraction for Li et al., “ATAD3 Is a Limiting Factor in Mitochondrial Biogenesis and Adipogenesis of White Adipocyte-Like 3T3-L1 Cells”

doi: 10.1128/mcb.01170-13

Figure Lengend Snippet: Figure 5 - Effects of ATAD3 and Drp1/Mfn2 overexpressions on lipogenesis and 984

Article Snippet: The sequences of double stranded RNAs for ATAD3 were: 229 Sequence 1: Forward 5’-ACAACAGCAACUUCUGAAUdTdT-3’, 230 Reverse 3’-dTdTUGUUGUCGUUGAAGACUUA-5’ 231 Sequence 2: Forward 5’-ACAGCAGUCCAAGCUCAAGdTdT-3’ 232 Reverse 3’-dTdTUGUCGUCAGGUUCGAGUUC-5’ 233 Plasmid encoding mouse ATAD3 is pCDNA3.1+ based; Drp1- and Mfn2-Myc-encoding 234 plasmids are respectively P26048 and P23213 pCDNA3-based from Addgene.

Techniques: