Journal: Biomolecules

Article Title: Exogenous Selenoprotein V Induces Apoptosis in Murine Testicular Teratoma Cells via Mitochondrial Dysfunction and ROS Overproduction

doi: 10.3390/biom15121733

Figure Lengend Snippet: Effects of 24-h pre-incubation of F9 cells with protein solvent buffer or various concentrations of recombinant SELENOV on cell viability. ( A ) Viability cytogram of F9 cells following treatment with 100 μL/mL (equivalent to 50 μg/mL SELENOV application) or 200 μL/mL (equivalent to 100 μg/mL SELENOV application) of solvent buffer. The 10 μg/mL SELENOV group was compared to the untreated control, as the corresponding buffer volume was negligible. ( B ) Viability cytogram of F9 cells after treatment with different concentrations of recombinant SELENOV. ( C ) Effects of solvent buffer or recombinant SELENOV on cell viability and activation of cell death pathways. Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. *** p < 0.001 versus the Control group; ### p < 0.001 versus the corresponding Buffer group.

Article Snippet: The study utilized several established cell lines: F9 cells (a murine embryonal carcinoma model derived from testicular teratoma of C57BL/6 mice) and mouse fibroblasts (L-929), all obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Incubation, Solvent, Recombinant, Control, Activation Assay

Journal: BMC Pharmacology & Toxicology

Article Title: Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

doi: 10.1186/s40360-017-0151-8

Figure Lengend Snippet: Development of neurons derived from RA-treated P19 and SH-SY5Y cells, and NGF-stimulated PC12 cells up to 10 days in culture. The cells were plated at a density of 500 cells/mm 2 and immunostained against the neuron-specific protein βIII-tubulin. a Representative fluorescence microscopy images of neurons (20 × magnification). b Fluorescence of anti-βIII-tubulin antibodies measured in a microplate reader and expressed as relative fluorescence units (RFU). Data are means ± SEM of 3–4 independent experiments

Article Snippet: P19 mouse embryonal carcinoma cells (cat. no. ECACC 95102107), PC12 rat adrenal pheochromocytoma cells (cat. no. ECACC 88022401) and SH-SY5Y human neuroblastoma cells (cat. no. ECACC 94030304) were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Porton Down, UK).

Techniques: Derivative Assay, Fluorescence, Microscopy

Journal: BMC Pharmacology & Toxicology

Article Title: Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

doi: 10.1186/s40360-017-0151-8

Figure Lengend Snippet: Concentration-dependent effects of MeHg, okadaic acid and acrylamide on cell viability and expression of the neuron-specific protein βIII-tubulin in neuronally differentiated P19, PC12 and SH-SY5Y cells. The cells were plated at a density of 500 cells/mm 2 and cultured for 6 days in the differentiation media, followed by exposure to the test compounds for 48 h. Effects of MeHg ( a , b ), okadaic acid ( c , d ) and acrylamide ( e , f ) on the cell viability were assessed by using the calcein-AM assay ( a , c , e ), and the immunofluorescence of βIII-tubulin ( b , d , f ). The data are means ± SEM of n = 6 independent experiments ( n = 3 for okadaic acid in the βIII-tubulin assay; panel d ). The results are expressed as percentage of non-treated cells or cells treated with 0.1% DMSO (used as vehicle). Wells treated with 2% Triton X-100 for 30 min served as controls for maximal cell death. Statistical analysis was performed using repeated measures one-way ANOVA with post hoc Dunnett’s multiple comparisons test (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001) compared to corresponding controls

Article Snippet: P19 mouse embryonal carcinoma cells (cat. no. ECACC 95102107), PC12 rat adrenal pheochromocytoma cells (cat. no. ECACC 88022401) and SH-SY5Y human neuroblastoma cells (cat. no. ECACC 94030304) were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Porton Down, UK).

Techniques: Concentration Assay, Expressing, Cell Culture, Calcein AM Assay, Immunofluorescence

Journal: BMC Pharmacology & Toxicology

Article Title: Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

doi: 10.1186/s40360-017-0151-8

Figure Lengend Snippet: Representative fluorescence microscopy images of neuronally differentiated P19, PC12 and SH-SY5Y cells exposed to 1 μM methylmercury, 10 nM okadaic acid and 1 mM acrylamide. The cells were plated at a density of 500 cells/mm 2 and cultured for 6 days in the differentiation media, followed by exposure to the test compounds for 48 h. The cells were immunolabeled against the neuron-specific protein βIII-tubulin and the fluorescence microscopy images were obtained at 20 × magnification

Article Snippet: P19 mouse embryonal carcinoma cells (cat. no. ECACC 95102107), PC12 rat adrenal pheochromocytoma cells (cat. no. ECACC 88022401) and SH-SY5Y human neuroblastoma cells (cat. no. ECACC 94030304) were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Porton Down, UK).

Techniques: Fluorescence, Microscopy, Cell Culture, Immunolabeling

Journal: BMC Pharmacology & Toxicology

Article Title: Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

doi: 10.1186/s40360-017-0151-8

Figure Lengend Snippet: Effects of MeHg, BSO and GSH on the viability of RA-treated P19 cells ( a ) and SH-SY5Y cells ( b ). Cells cultured for 6 days in the differentiation media were pre-treated with 100 μM BSO for 17 h or 1 mM GSH for 1 h followed by exposure to 1 μM MeHg for 24 h. Cell viability was assessed with the PrestoBlue assay that measures cellular metabolic reduction, and extracellular LDH activity assay. Data are means ± SEM of n = 6 independent experiments. For the PrestoBlue assay, data are expressed as percentage of non-treated or 0.1% DMSO vehicle-treated cells. For the LDH assay, the data are presented as percentage of total cell death (cells treated with 2% Triton X-100). Statistical analysis was undertaken using one-way ANOVA with post hoc Bonferroni’s multiple comparisons test: * p < 0.05, ** p < 0.01, and **** p < 0.0001 (the effect of the treatment compared to the corresponding vehicle control, or the effect of the combination of MeHg + GSH or MeHg + BSO compared to MeHg per se ). ND = not determined

Article Snippet: P19 mouse embryonal carcinoma cells (cat. no. ECACC 95102107), PC12 rat adrenal pheochromocytoma cells (cat. no. ECACC 88022401) and SH-SY5Y human neuroblastoma cells (cat. no. ECACC 94030304) were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Porton Down, UK).

Techniques: Cell Culture, Prestoblue Assay, Activity Assay, Lactate Dehydrogenase Assay, Control

Journal: BMC Pharmacology & Toxicology

Article Title: Comparison of neurons derived from mouse P19, rat PC12 and human SH-SY5Y cells in the assessment of chemical- and toxin-induced neurotoxicity

doi: 10.1186/s40360-017-0151-8

Figure Lengend Snippet: The effects of MeHg, GSH and BSO on TMRE fluorescence in RA-treated P19 ( a ) and SH-SY5Y cells ( b ). Cells cultured for 6 days in the differentiation media were pre-treated with 100 μM BSO for 17 h or with 1 mM GSH for 1 h followed by exposure to 1 μM MeHg for 24 h. Alterations in mitochondrial membrane potential were measured with the TMRE assay. Data are means ± SEM of n = 6 independent experiments, and expressed as percentage of non-treated or 0.1% DMSO vehicle-treated cells. Statistical analysis was undertaken using one-way ANOVA with post hoc Bonferroni’s multiple comparisons test: * p <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 (the effect of the treatment compared to the corresponding vehicle control, or the effect of the combination of MeHg + GSH or MeHg + BSO compared to MeHg per se )

Article Snippet: P19 mouse embryonal carcinoma cells (cat. no. ECACC 95102107), PC12 rat adrenal pheochromocytoma cells (cat. no. ECACC 88022401) and SH-SY5Y human neuroblastoma cells (cat. no. ECACC 94030304) were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Porton Down, UK).

Techniques: Fluorescence, Cell Culture, Membrane, Control