ABSTRACT: Gene Expression Profile of Medullary Epithelial Cells isolated from thymus of Bone Marrow (BM) reconstituted RAG1 null (control) and SCID NOD mouse
Project description: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
Project description: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.
Project description:OCI-AML3, a human acute myeloid leukemia cell line, was inoculated into NOD/SCID/IL-2rγnull (NSG) mice. After engraftment and leukemic expansion were documented, mice were divided into 3 groups: control, or treatment for 24 or 72 hours with daily injections of LY2510294, a CXCR4-inhibitory peptide. OCI-AML3 cells were isolated from peripheral blood (PB), bone marrow (BM), or spleen (SP) for gene expression profiling (GEP).
Project description:compare the gene expression profile between irradiated Lin-Sca-1+c-Kit+ (LSK) cells from mouse bone marrow reconstituted with wild type and necdin null fetal liver cells The Affymetrix oligonucleotide array was used for this analysis compare the gene expression profile betweenirradiated Lin-Sca-1+c-Kit+ (LSK) cells from mouse bone marrow reconstituted with wild type and necdin null fetal liver cells
Project description:The oligo microarrays were used to determine mRNA expression profiles of medullary thymic epithelial cells (mTECs) isolated from thymus of pre-diabetic NOD mice.
Project description:To investigate how HSCs functionally compensate for the B cell deficiencies of other HSCs within an organism, we co-transplanted wildtype (WT) HSCs and lineage-deficient HSCs into lethally irradiated WT recipient mice. WT HSCs were then purified from the bone marrow of recipient mice for RNA isolation and sequencing. The purpose is to determine whether there are common genes shared between compensating WT HSCs in the WT co-transplanted with uMT-/- (B6.129S2(B6)-Ighmtm1Cgn/J) and WT co-transplanted with NSG (NOD-scid IL2Rgamma null).
Project description:Due to high sequence identity between human and M. musculus microRNAs, we used in this study human microRNA microarrays to determine microRNA expression profiles of medullary thymic epithelial cells (mTECs) from thymus of pre-diabetic NOD mice.
Project description:compare the gene expression profile between irradiated Lin-Sca-1+c-Kit+ (LSK) cells from mouse bone marrow reconstituted with wild type and necdin null fetal liver cells The Affymetrix oligonucleotide array was used for this analysis
Project description:In order to identify genes expressed by cells that leave the spleen, the spleens were harvested from untreated reconstituted humanized mice (N=4) and a single cell suspension was prepared . The cultures were treated with either teplizumab (anti human CD3 hOKT3g1(Ala-Ala)) or hIg for 18 hrs in vitro. Splenocytes were also harvested 18 hrs after reconstituted mice (N=4) were treated with teplizumab in vivo. The humanized mice used where NOD/SCID IL2gc-/- (NSG) reconstituted with human CD34+ at birth.
