Journal: Computational and Structural Biotechnology Journal

Article Title: A Review on Genomics APIs

doi: 10.1016/j.csbj.2015.10.004

Figure Lengend Snippet: Workflow for the 23andMe API. A 23andMe customer, through a third-party application, sends API calls requesting for their genotypes within a given chromosomal location. This information is present within the 23andMe database as the customer's genetic test analysis report. Once the third-party application is successfully authenticated, the appropriate response is sent back to the customer. This response can then be combined with other sources of information for a more detailed downstream analysis and visualization.

Article Snippet: Unlike the other two APIs, there is really no easy way to create a reference implementation for the 23andMe API since the data are securely managed within 23andMe's database and the only people with access to it are personnel within that organization.

Techniques:

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effects of resveratrol, quercetin, apigenin and 7-ketocholesterol on cell viability and cell growth determined with the fluorescein diacetate and sulforhodamine 101 assays. Murine neuro-2a (N2a) neuroblastoma cells were cultured for 48 h with or without resveratrol (RSV), quercetin (QCT), apigenin (API) or 7-ketocholesterol (7KC; 1.5625 to 100 µM). The results are in percentages relative to the control (untreated cells). Data obtained with the fluorescein diacetate (FDA) assay ( A – D ), and the sulforhodamine (SR101) assay ( E – H ) are shown. Data shown are expressed as mean ± standard deviation (SD) of four independent experiments performed in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API- and 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less.

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Cell Culture, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effects of resveratrol, quercetin, apigenin or α-tocopherol on 7-ketocholesterol-induced plasma membrane permeability evaluated with propidium iodide. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as the positive control for cytoprotection, or with polyphenols: resveratrol (RSV) ( A ), quercetin (QCT) ( B ) or apigenin (API) ( C ) at concentrations ranging from 1.5625 (written 1.5) to 26 µM. Plasma membrane permeability was measured by flow cytometry with propidium iodide (PI): for each assay, the percentage of PI positive cells was determined. Two vehicle controls were realized: Ethanol (EtOH) (0.2%) used with RSV and 7KC, and DMSO (0.2%) used with QCT and API. Each value is the mean ± standard deviation (SD) of four independent experiments. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Permeability, Cell Culture, Positive Control, Flow Cytometry, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin and α-tocopherol on 7-ketocholesterol-induced oxidative stress: ROS overproduction and catalase activity, measurement of superoxide dismutase and glutathione peroxidase activity. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at a concentration of 3.125 and/or 6.25 µM. ( A ) ROS overproduction was measured by flow cytometry after staining with dihydroethidine (DHE) and evaluated by the percentage of DHE positive cells. The effect on catalase activity, a peroxisomal antioxidant enzyme, which degrades hydrogen peroxide (H 2 O 2 ), was determined by a colorimetric assay with the measurement of H 2 O 2 consumption. The enzymatic activities of superoxide dismutase (SOD) ( B ) and glutathione peroxidase (GPx) ( C ) were determined by colorimetric assays. Data shown are mean ± standard deviation (SD) of three independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco)- treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Activity Assay, Cell Culture, Positive Control, Concentration Assay, Flow Cytometry, Staining, Colorimetric Assay, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin or α-tocopherol on 7KC-induced oxidative stress: measurement of GPx1 , Nrf2 , Sod1 and Sod2 mRNA levels by real time-quantitative polymerase chain reaction. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at a concentration of 3.125 µM. The relative expression of GPx1 ( A ), Nrf2 ( B ), Sod1 ( C ) and Sod2 ( D ) mRNAs was determined by real time-quantitative polymerase chain reaction (RT-qPCR). In untreated cells, the cycle threshold (Ct) values are provided for each gene studied: GPx1 (Ct = 24 ± 1), Nrf2 (Ct = 21 ± 1), Sod1 (Ct = 20± 1) and Sod2 (Ct = 23 ± 1). Data shown are mean ± standard deviation (SD) of two independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Positive Control, Concentration Assay, Expressing, Quantitative RT-PCR, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effects of resveratrol, quercetin, apigenin, α-tocopherol and 7-ketocholesterol on the expression of antioxidant proteins (Nrf2, GPx1, SOD1, SOD2, catalase), mitochondrial biogenesis proteins (AMPKα, SIRT1, PGC-1α) and enzymes involved in the peroxisomal β-oxidation (ABCD1, MFP2) in murine N2a neuroblastoma cells. N2a cells previously cultured for 24 h were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) at 3.125 µM or α-tocopherol (400 µM). Representative Western blots for analysis were presented, showing protein abundance of antioxidant protein markers (expression of nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase (GPx1), superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2) and catalase), mitochondrial biogenesis proteins (AMP-activated protein kinase- alpha (AMPKα), Sirtuin 1 (SIRT1) and peroxisome proliferator-activated receptor gamma co-activator-1 alpha (PGC-1α)) and of proteins involved in peroxisomal β-oxidation (ATP binding cassette subfamily D member 1 (ABCD1) and peroxisomal multifunctional enzyme type 2 (MFP2). β-actin was used as the loading control. The intensities of the bands for each set were individually determined and are presented as the ratio over β-actin signal. The EtOH value (0.2%) and the DMSO value (0.2%) correspond to the final EtOH and DMSO concentration in the culture medium. No difference was observed between control and vehicle (EtOH and DMSO)-treated cells. Data shown are representative of three independent experiments. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Expressing, Cell Culture, Western Blot, Binding Assay, Concentration Assay, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effects of resveratrol, quercetin, apigenin and α-tocopherol on 7-ketocholesterol-induced mitochondrial dysfunction: loss of mitochondrial membrane potential measured with DiOC 6 (3) and mitochondrial ROS overproduction measured with MitoSOX. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at concentrations of 3.125 and 6.25 µM. Loss of mitochondrial membrane potential (ΔΨm) was measured by flow cytometry after staining with DiOC 6 (3) and evaluated by the percentage of DiOC 6 (3) negative cells ( A ). The effect on ROS overproduction at the mitochondrial level was determined by flow cytometry after staining with MitoSOX and evaluated by the percentage of MitoSOX positive cells ( B ). Data shown are mean ± standard deviation (SD) of three independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Cell Culture, Positive Control, Flow Cytometry, Staining, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin or α-tocopherol on 7KC-induced mitochondrial biogenesis disorders: measurement of the mRNA levels of Ampkα1, Sirt1 and Pgc-1α by real time-quantitative polymerase chain reaction. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetine (QCT) or apigenin (API) used at a concentration of 3.125 µM. The relative expression of Ampkα1 ( A ), Sirt1 ( B ), and Pgc-1α ( C ) mRNAs was determined by real time-quantitative polymerase chain reaction (RT-qPCR). In untreated cells, the cycle threshold (Ct) values are provided for each gene studied: Ampkα1 (Ct = 29 ± 1), Sirt1 (Ct = 26 ± 1), and Pgc-1α (Ct = 28 ± 1). Data shown are mean ± standard deviation (SD) of two independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Positive Control, Concentration Assay, Expressing, Quantitative RT-PCR, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Flow cytometric evaluation of the effect of resveratrol, quercetin, apigenin or α-tocopherol on 7KC-induced changes of peroxisomal mass evaluated with ABCD3 expression. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at a concentration of 3.125 µM. The level of ABCD3 was evaluated by flow cytometry: the percentage of cells with lower ABCD3 levels, compared to untreated cells and vehicle-treated cells (EtOH 0.2%; DMSO 0.2%) was determined. Data shown are mean ± standard deviation (SD) of three independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Expressing, Cell Culture, Positive Control, Concentration Assay, Flow Cytometry, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin or α-tocopherol on 7KC-induced changes of peroxisomal mass and biogenesis: measurement of the mRNA levels of Abcd3 , Pex13 and Pex14 by real time-quantitative polymerase chain reaction. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at a concentration of 3.125 µM. The relative expression of Abcd3 ( A ), Pex13 ( B ), and Pex14 ( C ) mRNAs was determined by real time-quantitative polymerase chain reaction (RT-qPCR). In untreated cells, the cycle threshold (Ct) values are provided for each gene studied: Abcd3 (Ct = 21 ± 1), Pex13 (Ct = 24 ± 1), and Pex14 (Ct = 24 ± 1). Data shown are mean ± standard deviation (SD) of two independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Positive Control, Concentration Assay, Expressing, Quantitative RT-PCR, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin and α-tocopherol on 7KC-induced changes of peroxisomal β-oxidation: measurement of the mRNA levels of Abcd1 , Acox1 , Mfp2 and Thiolase A by real time-quantitative polymerase chain reaction. N2a cells were cultured for 48 h with or without 7-ketocholesterol (7KC, 50 μM) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetine (QCT) or apigenin (API) used at a concentration of 3.125 µM. Beta-Oxidation of very long chain fatty acids (VLCFAs) in peroxisomes: the implication of ABCD1, ACOX1, MFP2 and Thiolase A in peroxisomal β-oxidation is summarized in ( A ). The relative expression of Abcd1 ( B ), Acox1 ( C ), Mfp2 ( D ) and Thiolase A ( E ) mRNAs was determined by real time-quantitative polymerase chain reaction (RT-qPCR). In untreated cells, the cycle threshold (Ct) values are provided for each gene studied: Abcd1 (Ct = 22 ± 1), Acox1 (Ct = 23 ± 1), Mfp2 (Ct = 26 ± 1) and Thiolase A (Ct = 26 ± 1). Data shown are mean ± standard deviation (SD) of two independent experiments conducted in triplicate. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Positive Control, Concentration Assay, Expressing, Quantitative RT-PCR, Standard Deviation, MANN-WHITNEY

Journal: Cells

Article Title: Prevention by Dietary Polyphenols (Resveratrol, Quercetin, Apigenin) Against 7-Ketocholesterol-Induced Oxiapoptophagy in Neuronal N2a Cells: Potential Interest for the Treatment of Neurodegenerative and Age-Related Diseases

doi: 10.3390/cells9112346

Figure Lengend Snippet: Effect of resveratrol, quercetin, apigenin and α-tocopherol on 7KC-induced autophagy and apoptosis: characterization of apoptotic nuclei by staining with Hoechst 33342, identification of cleaved-caspase-3 and PARP and activation of LC3-I into LC3-II by Western blotting. N2a cells were cultured with or without 7-ketocholesterol (7KC: 50 μM, 48 h) in the presence or absence of α-tocopherol (α-toco: 400 µM) used as a positive control for cytoprotection, or with polyphenols: resveratrol (RSV), quercetin (QCT) or apigenin (API) used at 3.125 µM. ( A ) Cells with condensed nuclei (cn) and/or fragmented nuclei (fn) characteristic of apoptotic cells are identified. When 7KC was associated with RSV, QCT and API, the presence of apoptotic cells was strongly reduced; no or few apoptotic cells were present in control (untreated cells), vehicle-treated cells (EtOH 0.2%; DMSO 0.2%) and RSV-, QCT- and API-treated cells. ( B ) Apoptosis was also evaluated by caspase-3 activation (cleaved caspase-3), and PARP fragmentation, and autophagy was evaluated by conversion of LC3-I to LC3-II (increased ratio (LC3-II / LC3-I)). The EtOH value (0.2%) and the DMSO value (0.2%) correspond to the final EtOH and DMSO concentration in the culture medium. No difference was observed between control and vehicle (EtOH and DMSO)-treated cells. Data shown are representative of three independent experiments. Significance of the differences between control (untreated cells) and RSV-, QCT-, API-, α-toco- or 7KC-treated cells; Mann–Whitney test: * p < 0.05 or less. Significance of the differences between 7KC-treated cells and (7KC + (RSV, QCT, API or α-toco))-treated cells; Mann–Whitney test: # p < 0.05 or less. No significant differences were found between control and vehicle-treated cells (Ethanol (EtOH): 0.2% and DMSO: 0.2%).

Article Snippet: 7KC (ref: C2394; purity >90%), RSV (trans-resveratrol, ref: 501-36-0; purity 99%) and QCT (ref: #Q0125; purity >98%) were from (Sigma-Aldrich, St Quentin-Fallavier, France); API was from (Euromedex, Souffelweyersheim, France; ref: #52262; purity >97%).

Techniques: Staining, Activation Assay, Western Blot, Cell Culture, Positive Control, Concentration Assay, MANN-WHITNEY