Journal: Cancers

Article Title: SIRT2 Contributes to the Resistance of Melanoma Cells to the Multikinase Inhibitor Dasatinib

doi: 10.3390/cancers11050673

Figure Lengend Snippet: Cellular WM853 and MDA-MB-435S clone characteristics. ( a ) SIRT2 expression in SCW3 and SSW30 clones of WM853 cells and SCM1 and SSM15 clones of MDA-MB-435S as evidenced by Western blotting. ( b ) The cytotoxic effects of dasatinib treatment (48 h) on melanoma SCW3 and SSW30 clones of the WM853 cell line, as evidenced using the neutral red assay, mean ± SD, ( n = 3, independent experiments) ( c ) The cytotoxic effects of dasatinib treatment (48 h) on melanoma SCM1 and SSM15 clones of the MDA-MB-435S cell line, as evidenced using the neutral red assay, mean ± SD, ( n = 3, independent experiments).

Article Snippet: The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich, St. Louis, MO, USA), 20% Leibovitz’s (Sigma-Aldrich, St. Louis, MO, USA), 2% fetal bovine serum (Pan Biotech GmbH, Aidenbach, Germany), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Clone Assay, Western Blot, Neutral Red Assay

Journal: Cancers

Article Title: SIRT2 Contributes to the Resistance of Melanoma Cells to the Multikinase Inhibitor Dasatinib

doi: 10.3390/cancers11050673

Figure Lengend Snippet: Selected category of genes (based on a literature search) with changes in expression in WM853 cells with SIRT2 downregulation as determined using RNA-seq.

Article Snippet: The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich, St. Louis, MO, USA), 20% Leibovitz’s (Sigma-Aldrich, St. Louis, MO, USA), 2% fetal bovine serum (Pan Biotech GmbH, Aidenbach, Germany), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing

Journal: Cancers

Article Title: SIRT2 Contributes to the Resistance of Melanoma Cells to the Multikinase Inhibitor Dasatinib

doi: 10.3390/cancers11050673

Figure Lengend Snippet: Treatment with dasatinib impairs the migration of melanoma cells based on the scratch assay. ( a ) Image from a single representative experiment performed using WM853 SCW3 and SSW30 clones. ( b ) Image from a single representative experiment performed using MDA-MB-435S SCM1 and SSM15 clones. Quantification of above results is shown in the . 10 × 10 magnification.

Article Snippet: The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich, St. Louis, MO, USA), 20% Leibovitz’s (Sigma-Aldrich, St. Louis, MO, USA), 2% fetal bovine serum (Pan Biotech GmbH, Aidenbach, Germany), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Migration, Wound Healing Assay, Clone Assay

Journal: Cancers

Article Title: SIRT2 Contributes to the Resistance of Melanoma Cells to the Multikinase Inhibitor Dasatinib

doi: 10.3390/cancers11050673

Figure Lengend Snippet: SIRT2 downregulation impairs the response of melanoma cells to EGFR and EPHA2 activators. The generated melanoma clones were treated with selected concentrations of EGF (EGFR activator) and ephrin A1 (EPHA2 activator) for 10 min and/or 30 min, respectively. Then, protein lysates were prepared and analyzed by Western blotting. ( a ) Experiments performed using WM853 SCW3 and SSW30 clones. ( b ) Experiments performed using MDA-MB-435S SCM1 and SSM15 clones.

Article Snippet: The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich, St. Louis, MO, USA), 20% Leibovitz’s (Sigma-Aldrich, St. Louis, MO, USA), 2% fetal bovine serum (Pan Biotech GmbH, Aidenbach, Germany), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Generated, Clone Assay, Western Blot

Journal: Cancers

Article Title: SIRT2 Contributes to the Resistance of Melanoma Cells to the Multikinase Inhibitor Dasatinib

doi: 10.3390/cancers11050673

Figure Lengend Snippet: SIRT2 downregulation impairs the expression and phosphorylation of tyrosine kinase receptor-associated pathways. The generated melanoma clones were treated with selected concentrations of dasatinib for 1 h. Then, protein lysates were prepared and analyzed by Western blotting. ( b ) Experiments performed using WM853 SCW3 and SSW30 clones. ( a ) Experiments performed using MDA-MB-435S SCM1 and SSM15 clones.

Article Snippet: The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich, St. Louis, MO, USA), 20% Leibovitz’s (Sigma-Aldrich, St. Louis, MO, USA), 2% fetal bovine serum (Pan Biotech GmbH, Aidenbach, Germany), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Phospho-proteomics, Generated, Clone Assay, Western Blot

Journal: Oncogene

Article Title: Pyruvate dehydrogenase inactivation causes glycolytic phenotype in BAP1 mutant uveal melanoma

doi: 10.1038/s41388-021-02154-0

Figure Lengend Snippet: a. Average expression of PDH (E1), DLAT (E2) and PDHK1 between BAP1 wild type (92.1, OMM1.3, UM001, UM004, UM002B and MM66) and mutant (MP46, MP65, MP38, MM28, PDX4 and WM3618F) UM cells were analyzed by RPPA analysis. RPPA data were used to determine antibodies that are significantly different between mutant and wild-type cell lines. Comparison of average normalized log2 values were performed by the two-sample t-test method and assumed unequal variance. Data points are shown as individual lysate collections **p<0.01. b. Validation of protein expression by Western blot.

Article Snippet: WM3618F was purchased from Rockland Immunochemicals (Limerick, PA) and cultured routinely in 1× MCDB media containing 10% heat-inactivated FBS, CaCl 2 , 50 IU penicillin and 50 μg/mL streptomycin.

Techniques: Expressing, Mutagenesis, Western Blot