Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Effect of supercritical CO 2 tomato extract (sCO 2 TE) on cell viability. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or sodium arsenite (AsNaO 2 , 200 μM). The number of viable cells was determined by Trypan Blue (TB) exclusion test. Data are reported as the mean ± standard deviation (SD) among 10 independent experiments ( A ). Cell proliferation was measured by 3-(4,5-dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data are reported as the mean ± standard deviation (SD) among 5 independent experiments performed in triplicate and represent cell viability as a percentage of vehicle ( B ).

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Standard Deviation, MTT Assay

Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Effect of supercritical CO 2 tomato extract (sCO 2 TE) on DNA. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or sodium arsenite (AsNaO 2 , 200 μM) and, after staining with propidium iodide (PI), were analyzed by flow cytometric analysis to evaluate cell cycle phases. Data are reported as the mean ± standard deviation (SD) among 3 independent experiments.

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Staining, Standard Deviation

Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Analysis of intracellular reactive oxygen species (ROS) after supercritical CO 2 tomato extract (sCO 2 TE) treatment. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or sodium arsenite (AsNaO 2 , 200 μM), and alternatively they were pre-treated with sCO 2 TE (100 μg/mL) and then stimulated with AsNaO 2 (200 μM). After treatments, ROS production was assessed using the DCFH-DA fluorescent probe. Flow cytometric histograms show measurement of ROS levels through flow cytometry analysis ( A ). Quantified ROS levels: histograms show the mean fluorescence intensity (MFI) ( B ). The results are presented as mean ± SD from 3 independent experiments. Statistical analysis indicates: * p < 0.05 and *** p < 0.001.

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Flow Cytometry, Fluorescence

Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Analysis of lipid peroxidation after supercritical CO 2 tomato extract (sCO 2 TE) treatment. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or sodium arsenite (AsNaO 2 , 200 μM), and alternatively they were pre-treated with sCO 2 TE (100 μg/mL) and then stimulated with AsNaO 2 (200 μM). After treatments, lipid peroxidation was evaluated using BODIPY 581/591 C11probe (Lipid Peroxidation Sensor that results in a FITC emission). Flow cytometric histograms show measurement of lipid peroxidation through flow cytometry analysis ( A ). Quantified lipid peroxidation, histograms show the mean fluorescence intensity (MFI) ( B ). The results are presented as mean ± SD from 3 independent experiments. Statistical analysis indicates: **** p < 0.0001.

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Flow Cytometry, Fluorescence

Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Analysis of NRF-2 phosphorylation after supercritical CO 2 tomato extract (sCO 2 TE) treatment. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or sodium arsenite (AsNaO 2 , 200 μM), and alternatively they were pre-treated with sCO 2 TE (100 μg/mL) and then stimulated with AsNaO 2 (200 μM). After treatments, cells were lysed and analyzed by Western blot. Nuclear cell extracts were analyzed to verify phosphorylated NRF2 using rabbit anti-phospho-NRF2 Ab. As a control, for loading and purity of preparation, membrane was stripped and reprobed with polyclonal anti-HISTONE H1 Ab. Densitometric phospho-NRF2/HISTONE H1 ratios are shown in the right panel. Results represent the mean ± SD from 3 independent experiments. Statistical analysis indicates: ** p < 0.01; **** p < 0.0001.

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Phospho-proteomics, Western Blot, Control, Membrane

Journal: Nutrients

Article Title: Supercritical CO 2 -Derived Tomato Extract Activates Signaling Pathways to Reduce Oxidative Stress and Inflammation in Astrocyte Cells

doi: 10.3390/nu18091464

Figure Lengend Snippet: Analysis of MAPK signaling and NF-κB activation after supercritical CO 2 tomato extract (sCO 2 TE) treatment. Human glioblastoma astrocyte cells U-373 were treated with sCO 2 TE (100 μg/mL), conventional tomato extract (CTE, 100 μg/mL) or LPS (100 ng/mL), and alternatively they were pre-treated with sCO 2 TE (100 μg/mL) and then stimulated with LPS (100 ng/mL). After treatments cells were lysed and analyzed by Western blot. ( A ) Whole cell extracts were analyzed to verify phosphorylated-ERK expression using rabbit anti-phospho-ERK1/2 Ab. ( B ) Nuclear cell extracts were analyzed to verify phosphorylated-NF-kB-p65 expression using rabbit anti-phospho-NF-kB-p65 Ab. As a control, for loading and purity of preparation, membrane was stripped and reprobed with polyclonal anti-HISTONE H1 Ab. Densitometric phospho-ERK/total ERK and phospho-NF-kB-p65/HISTONE H1 ratios are shown in the right panels. Results represent the mean ± SD from 3 independent experiments. Statistical analysis indicates: ** p < 0.01; **** p < 0.0001.

Article Snippet: Human glioblastoma astrocytoma cells U-373 MG (American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in DMEM F-12 medium (Sigma-Aldrich, Milan, Italy) supplemented with 10% fetal bovine serum (FBS, Aurogene S.r.l., Rome, Italy), 100 units/mL penicillin and 10 mg/mL streptomycin (Aurogene S.r.l.) and maintained in a humified atmosphere of 37 °C and 5% CO 2 .

Techniques: Activation Assay, Western Blot, Expressing, Control, Membrane