Journal: Cancers
Article Title: A Novel Cyclic Pentadepsipeptide, N -Methylsansalvamide, Suppresses Angiogenic Responses and Exhibits Antitumor Efficacy against Bladder Cancer
doi: 10.3390/cancers13020191
Figure Lengend Snippet: MSSV-mediated inhibition of proliferation of the human bladder cancer cell lines, 5637 and T24, was owing to G1-phase cell cycle arrest. ( A ) Both cell lines were treated with or without MSSV at the indicated concentrations for 24 h, followed by MTT assays for cell viability. ( B ) Cells were treated with indicated concentrations of MSSV for 24 h, and cell counting was performed via trypan blue staining. ( C ) Both cell lines were treated with MSSV for 24 h and analyzed for cell cycle distribution using fluorescence-activated cell sorting (FACS) histograms. The percentage of cells in each cell cycle phase is presented. ( D ) Cells were exposed to MSSV at the indicated concentrations for 24 h. The expression levels of cyclin D1, cyclin E, CDK2, CDK4, p21WAF1, p27KIP1, p53, and GAPDH were analyzed via immunoblotting. Bar graphs show the relative fold changes in proteins at different MSSV concentrations in comparison with the control. ( E ) The cell lysates were immunoprecipitated with antibodies recognizing CDK2 and CDK4, followed by immunoblotting with specific antibodies against p21WAF1, p27KIP1, CDK2, and CDK4. Graphs show the relative amount of immunoprecipitated proteins as fold changes in comparison with the control. For the bar graphs, values were presented as the mean ± SD of three independent experiments; * p < 0.05, compared with the control group. Uncropped Western Blot Images in .
Article Snippet: The human bladder carcinoma cell lines, 5637 and T24, were purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a 5% CO 2 humidified incubator.
Techniques: Inhibition, Cell Counting, Staining, Fluorescence, FACS, Expressing, Western Blot, Comparison, Control, Immunoprecipitation