Article Title: A novel mouse model of rhabdomyosarcoma underscores the dichotomy of MDM2-ALT1 function in vivo
Figure Lengend Snippet: Transgenic mice expressing MDM2-ALT1 in B cells show significantly reduced populations of cells with B cell markers and defects in proliferation in spleens compared to controls A. Spleens of age-matched control ( MDM2-ALT1 −/− ; CD19-Cre +/− ; p53 +/+ ) and experimental ( 2C12-MDM2-ALT1 +/− ; CD19-Cre +/− ; p53 +/+ ) mice were harvested at 18 months and the splenocytes were stained for B cell markers CD19, B220, IgG or IgM or T cell markers CD3e or CD5. Compared to MDM2-ALT1-negative control mice ( n = 18) the MDM2-ALT1-positive experimental ( n = 15) mice showed a statistically significant decrease in the population of cells expressing B cell markers but no changes in T cell population. B. Splenocytes isolated from control ( n = 4) and experimental ( n = 5) mice were labeled with a fluorescence marker, CFSE, and stimulated for 72 hours with lipopolysaccharides (LPS), CD40 ligand, or anti-IgM molecules. When stimulated with CD40, the MDM2-ALT1 expressing experimental cohort showed a significantly lower proliferative response compared to splenocytes from control mice. The proliferating population was measured by gating for cells displaying low CFSE fluorescence intensity, indicative of dilution of the dye upon cell division. The percent proliferating population from each group was normalized to the respective non-stimulated control set (No stim). * indicates p
Article Snippet: Cells were stained for 30 minutes at 4°C with a combination of either anti-B220, anti-CD5, anti-IgM and anti-CD19 or anti-IgG, anti-CD3e, anti-IgM and anti-CD19 antibodies, then stained with PE-p53, FITC-Bcl-2, FITC-Annexin V (Life Technologies) or fixed and stained with propidium iodide.
Techniques: Transgenic Assay, Mouse Assay, Expressing, Staining, Negative Control, Isolation, Labeling, Fluorescence, Marker