The Nb2 lymphoma cell line is susceptible to glucocorticoid (GC)-induced apoptosis in the absence of mitogen and this effect is inhibited by the mitogen prolactin (PRL). This process is believed to be mediated by the glucocorticoid receptor (GR) and various downstream signals. The purpose of this study was to use the limiting dilution method to isolate subsets of Nb2 clones which expressed either highly enriched amounts of GR or an absence of GR and to relate the amount of GR to the magnitude of GC induced cytolysis. The presence of GR in these clones was determined by immunocytochemistry (ICC) using an affinity purified polyclonal rabbit antibody directed towards the GR. The presence of GR was then correlated to cytolysis in these cells produced by incubation with dexamethasone (Dex), a synthetic GC, in the presence and absence of PRL. Cytolysis was measured by two different methods, Trypan Blue exclusion counts, to determine the percentage of dead and the TUNEL (TdT-dependent dUTP-biotin Nick End Labeling) assay used to label apoptotic cells. The limiting dilution produced 28 clones with varying amounts of GR and all were responsive to Dex and this response was inhibited by PRL. A significant correlation (r2=0.419 and p
| Identifer | oai:union.ndltd.org:vcu.edu/oai:scholarscompass.vcu.edu:etd-6330 |
| Date | 01 January 1999 |
| Creators | Patel, Devang M. |
| Publisher | VCU Scholars Compass |
| Source Sets | Virginia Commonwealth University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
| Rights | © The Author |
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