Active Ingredients: Doxycycline
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We have examined signaling pathways by phosphoprotein fluorescence-activated cell sorter FACS analysis and found that atorvastatin induces a specific phosphoprotein signature by shutting down specific signaling networks required to sustain MYC activation.
Transgenic cell lines Single-cell suspensions were generated from tumors of MYC-overexpressing mice by mechanical disruption.
Early passages of the cell lines were used for the experiments described.
Proliferation assay Cells were grown in their respective media requirements, and cultures were pulsed for 18 hours with 0.
Cells were harvested, and thymidine incorporation was measured. Results are shown as the mean of triplicate experiments with standard error of the mean SEM given as a percentage.
Flow cytometry Intracellular probes for active kinases were made by conjugating phosphospecific antibodies to the Alexa Fluor dye series as described, and were used in phosphoprotein-optimized conditions.
All centrifugation, staining, and flow cytometry processes were performed on ice with ice-cold buffers using pretitered antibodies at optimal antibody concentrations and fluorophore to protein FTP ratios. Alexa Fluor dye series 405, 430, 488, 546, 568, 647, and 700 were from Molecular Probes.
Immunoblots were stripped by incubating with stripping buffer 62. Cells were monitored during infection. Infection was confirmed by GFP expression, and cells were then treated with atorvastatin.
All samples were analyzed in triplicates and were quantified using a standard curve generated by serial dilution of cDNA pooled from nontreated and treated lymphoma cells.
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