Gene Expression Profile of Medullary Epithelial Cells isolated from thymus of Bone Marrow (BM) reconstituted RAG1 null (control) and SCID NOD mouse

We identified DNAPK as one of the major proteins that physically interact with Autoimmune regulator (Aire). To establish physiological significance of DNAPK in Aire-driven expression of PTA genes in MECs, we utilized BM-reconstituted SCID mice (which express non functional DNAPK in their MECs) and RAG1 null mouse as a control. MECs from these mutated/reconstituted mice were sorted by Flow cytomety and their expression profile was analyzed using Affymetrix chips. The analysis shows that Aire-dependent PTA genes are significantly underexpressed in reconstituted SCIDS vs. RAGs (Controls), suggesting the involvement of DNAPK in Aire-mediated gene expression. Keywords: gene mutation Since SCID mice lack mature thymocytes and as a result thymic stroma, we reconstituted these animals by BM transfer (IP injection into 2-4d pups) 6 hrs post-irradiation (200rad). Reconstituted thymi were analyzed 6-8w after transfer.