Journal: BMC Veterinary Research

Article Title: Mycoplasma synoviae elongation factor thermo stable is an adhesion-associated protein that enters cells by endocytosis and stimulates DF-1 cell proliferation

doi: 10.1186/s12917-024-04374-4

Figure Lengend Snippet: Colocalization of M. synoviae EF-Ts with caveolin-1 (Cav-1) and clathrin heavy chain (CLTC). DF-1 cells were incubated with M. synoviae EF-Ts for 2 h and then fixed and labeled with caveolin-1 or clathrin heavy chain (red), rEF-Ts (green), and nuclei (blue). The fluorescence intensity curve and Pearson's R-value were determined by ImageJ software

Article Snippet: Pitstop-2 (HY-115604, MCE) is an inhibitor of clathrin protein-mediated endocytosis, and simvastatin (HY-17502, MCE) is an inhibitor of caveolin protein pathway-mediated endocytosis.

Techniques: Incubation, Labeling, Fluorescence, Software

Journal: Cell Research

Article Title: RAB31 marks and controls an ESCRT-independent exosome pathway

doi: 10.1038/s41422-020-00409-1

Figure Lengend Snippet: a Left, immunofluorescence of EGFR-HA (red) and Flag-RAB31 Q65L (magenta) with CD63-GFP (green) in Flag-RAB31 Q65L stable HeLa cells stably expressing shNC (negative control), shFLOT1, shFLOT2 or shFLOT1 and shFLOT2 and transiently expressing EGFR-HA and CD63-GFP under serum starvation (SS). Right up panel, the ratio of entry of EGFR-HA into CD63-GFP-positive LE/MVE in shNC ( n = 9 fields), shFLOT1 and shFLOT2 ( n = 9 fields), shFLOT1 ( n = 12 fields), shFLOT2 ( n = 12 fields). Right low panel, the ratio of entry of Flag-RAB31 Q65L into CD63-GFP-positive LE/MVE in shNC ( n = 9 fields), shFLOT1 and shFLOT2 ( n = 9 fields), shFLOT1 ( n = 12 fields), shFLOT2 ( n = 12 fields). b Western blotting analyses of the concentrated conditional media from the indicated stable HeLa cells used in a . c Up panels, immunofluorescence of EGFR-HA (red) and Flag-RAB31 (magenta) with FLOT1-GFP (green) in the indicated stable HeLa cells transiently expressing EGFR-HA and FLOT1-GFP under SS. Low panel left, the ratio of co-localization of EGFR-HA with FLOT1-GFP-positive vesicle in Vector ( n = 7 fields), RAB31 WT ( n = 8 fields) and RAB31 Q65L ( n = 9 fields). Low panel right, the ratio of co-localization of Flag-RAB31 with FLOT1-GFP-positive vesicle in RAB31 WT ( n = 8 fields) and RAB31 Q65L ( n = 9 fields). d Up panels, immunofluorescence of FLOT1-HA (red) and Flag-RAB31 (magenta) with CD63-GFP (green) in the indicated stable HeLa cells transiently expressing FLOT1-HA and CD63-GFP under SS. Low panel left, the ratio of co-localization of FLOT1-HA with CD63-GFP-positive LE/MVE in Vector ( n = 7 fields), RAB31 WT ( n = 7 fields) and RAB31 Q65L ( n = 8 fields). Low panel right, the ratio of entry of FLOT1-HA into CD63-GFP-positive LE/MVE in Vector ( n = 7 fields), RAB31 WT ( n = 7 fields) and RAB31 Q65L ( n = 8 fields). e Left, immunofluorescence of EGFR-HA (red) and Flag-RAB31 Q65L (magenta) with CD63-GFP (green) in Flag-RAB31 Q65L stable HeLa cells transiently expressing EGFR-HA and CD63-GFP and treated with DMSO, 5 μM GW4869, 5 μM simvastatin or 10 μM lovastatin under SS. Right up panel, the ratio of entry of EGFR-HA into CD63-GFP-positive LE/MVE in DMSO ( n = 8 fields), GW4869 ( n = 11 fields), simvastatin ( n = 13 fields) and lovastatin ( n = 12 fields). Right low panel, the ratio of entry of Flag-RAB31 Q65L into CD63-GFP-positive LE/MVE in DMSO ( n = 8 fields), GW4869 ( n = 11 fields), simvastatin ( n = 13 fields) and lovastatin ( n = 12 fields). f Western blotting analyses of the concentrated conditional media from the indicated stable HeLa cells used in e . All data are means ± SD. Unpaired t -test was used to analyze the difference between the two groups. **** P < 0.0001, *** P < 0.001, NS, no statistical significance. Scale bars, 10 μm.

Article Snippet: Simvastatin (Selleck; S1792) and Lovastatin (Selleck; S2061) were suspended in DMSO at the concentration of 10 mM in stock.

Techniques: Immunofluorescence, Stable Transfection, Expressing, Negative Control, Western Blot, Plasmid Preparation

Journal: BMC Cancer

Article Title: Heme oxygenase is not involved in the anti-proliferative effects of statins on pancreatic cancer cells

doi: 10.1186/s12885-016-2343-9

Figure Lengend Snippet: Anti-proliferative effect of statins on pancreatic cancer cells. The effect of simvastatin, cerivastatin, fluvastatin, lovastatin and pravastatin on cell proliferation was measured using MTT test in a ) PA-TU-8902, b ) MiaPaCa-2, and c ) BxPC-3 after 48 h of treatment. Statin concentration = 12 μM * p < 0.01, ** p < 0.0001 vs. control cells. CON, control cells; SIM, simvastatin; CER, cerivastatin; FLU, fluvastatin; LOV, lovastatin; PRA, pravastatin

Article Snippet: Cerivastatin, pravastatin and fluvastatin were purchased from LKT Laboratories, Inc (USA), lovastatin and simvastatin from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Concentration Assay, Control

Journal: BMC Cancer

Article Title: Heme oxygenase is not involved in the anti-proliferative effects of statins on pancreatic cancer cells

doi: 10.1186/s12885-016-2343-9

Figure Lengend Snippet: The effect of statins on activity and expression of HMOX in pancreatic cancer cells a ) HMOX activity was measured in PA-TU-8902, MiaPaCa-2 and BxPC-3 pancreatic cancer cell lines after 12 h of statin treatment (12 μM). b HMOX activity after 12 h of treatment were measured in PA-TU-8902, and c ) HMOX1 mRNA after 4 h of treatment. Hemin (30 μM) and PGJ2 (5 μM) served as positive controls for HMOX1 induction ability. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control cells CON, control cells; SIM, simvastatin; CER, cerivastatin; FLU, fluvastatin; LOV, lovastatin; PRA, pravastatin; PGJ2, 15-deoxy-Δ-12,14-prostaglandin J2

Article Snippet: Cerivastatin, pravastatin and fluvastatin were purchased from LKT Laboratories, Inc (USA), lovastatin and simvastatin from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Activity Assay, Expressing, Control

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Simvastatin Inhibits L-Type Ca2+-Channel Activity Through Impairment of Mitochondrial Function.

doi: 10.1093/toxsci/kfz068

Figure Lengend Snippet: Fig. 1. Simvastatin depolarizes the inner mitochondrial membrane potential, Δψmit. Representative traces of Rh123 fluorescence for primary mouse β-cells recorded in response to conditions indicated. Arrows show the

Article Snippet: Simvastatin and pravastatin were obtained from Tocris Bioscience (Bristol, UK).

Techniques: Membrane, Fluorescence

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Simvastatin Inhibits L-Type Ca2+-Channel Activity Through Impairment of Mitochondrial Function.

doi: 10.1093/toxsci/kfz068

Figure Lengend Snippet: Fig. 2. Simvastatin inhibits mitochondrial function in murine beta-cells. A) Effect of 0.1% vol/vol DMSO (n =

Article Snippet: Simvastatin and pravastatin were obtained from Tocris Bioscience (Bristol, UK).

Techniques:

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Simvastatin Inhibits L-Type Ca2+-Channel Activity Through Impairment of Mitochondrial Function.

doi: 10.1093/toxsci/kfz068

Figure Lengend Snippet: Figure 3. Glucose elicits calcium influx, which is inhibited by simvastatin. A) Representative intracellular calcium, [Ca2+]i, traces for five MIN6 beta-cells clusters all recorded in the same field in response to the

Article Snippet: Simvastatin and pravastatin were obtained from Tocris Bioscience (Bristol, UK).

Techniques:

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Simvastatin Inhibits L-Type Ca2+-Channel Activity Through Impairment of Mitochondrial Function.

doi: 10.1093/toxsci/kfz068

Figure Lengend Snippet: Figure 4. Simvastatin reverses the metabolic stimulation of L-type calcium channel activity. A-C) Single

Article Snippet: Simvastatin and pravastatin were obtained from Tocris Bioscience (Bristol, UK).

Techniques: Activity Assay

Journal: Journal of molecular and cellular cardiology

Article Title: MitoPlex: A Targeted Multiple Reaction Monitoring Assay for Quantification of a Curated Set of Mitochondrial Proteins

doi: 10.1016/j.yjmcc.2020.03.011

Figure Lengend Snippet: (A) Protein abundance heatmap for all MitoPlex proteins detected in organs of mice treated for 10 days with either DMSO or 20 mg/kg simvastatin. Expression is displayed as row z-score, ranging from undetected / below lower limit of quantification (white) to highest in the dataset (dark orange). (B) Log2-fold changes in mitochondrial proteins in the tissues of simvastatin-treated mice (vs. vehicle) as reported by MitoPlex; values are means ±SD (n=4), unpaired t-test, *p<0.05. (C) Log2-fold changes in mitochondrial proteins and metabolites of the TCA cycle in the tissues of mice treated with simvastatin (vs. vehicle). Proteins are labeled in red and metabolites in blue.

Article Snippet: Mouse Simvastatin Treatment Eight sixteen-week old C57/BL6 mice were injected with either vehicle (DMSO) or 20mg/kg of simvastatin (Toronto Research Chemicals, S485000) once a day intraperitoneally for 10 days [ 61 , 62 ].

Techniques: Expressing, Labeling

Journal: Journal of molecular and cellular cardiology

Article Title: MitoPlex: A Targeted Multiple Reaction Monitoring Assay for Quantification of a Curated Set of Mitochondrial Proteins

doi: 10.1016/j.yjmcc.2020.03.011

Figure Lengend Snippet: (A) Phase contrast microcopy images representative of C2C12 myotubes 6 days post differentiation in the presence of DMSO, 2uM simvastatin, 5uM CoQ1, or both. (B) Respirometry trace and respiratory rates from C2C12 myotubes differentiated in the presence DMSO, 2uM simvastatin, 5uM CoQ1, or both. One-way analysis of variance (ANOVA) vs DMSO, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are means ±SD, n=3. (C) Immunoblot analysis of whole cell lysates with the OXPHOS antibody cocktail and quantification of the results, normalized to Ponceau staining. One-way analysis of variance (ANOVA) vs DMSO, *p<0.05, **p< 0.01, ***p<0.001, ****p<0.0001; representative immunoblot is shown, values are means ±SD, n=3). (D) Protein abundance heatmap for all MitoPlex proteins detected in C2C12 myotubes differentiated in the presence DMSO, 2uM simvastatin, 5uM CoQ1, or both. Expression is displayed as row z-score, ranging from undetected / below lower limit of quantification (white) to highest in the dataset (dark orange). (E) Comparison of Log2-fold changes in mitochondrial proteins as reported by MitoPlex; values are means ±SD (n=3), unpaired t-test, *p<0.05 **p<0.01. (F) Heat map analysis of LC-MS metabolomics of samples from (D).

Article Snippet: Mouse Simvastatin Treatment Eight sixteen-week old C57/BL6 mice were injected with either vehicle (DMSO) or 20mg/kg of simvastatin (Toronto Research Chemicals, S485000) once a day intraperitoneally for 10 days [ 61 , 62 ].

Techniques: Western Blot, Staining, Expressing, Liquid Chromatography with Mass Spectroscopy

Journal: Journal of molecular and cellular cardiology

Article Title: MitoPlex: A Targeted Multiple Reaction Monitoring Assay for Quantification of a Curated Set of Mitochondrial Proteins

doi: 10.1016/j.yjmcc.2020.03.011

Figure Lengend Snippet: Log2-fold changes in mitochondrial proteins and metabolites of the TCA cycle in the tissues of mice treated with simvastatin (vs. vehicle). Proteins are labeled in red and metabolites in blue.

Article Snippet: Mouse Simvastatin Treatment Eight sixteen-week old C57/BL6 mice were injected with either vehicle (DMSO) or 20mg/kg of simvastatin (Toronto Research Chemicals, S485000) once a day intraperitoneally for 10 days [ 61 , 62 ].

Techniques: Labeling