title: Illumina HiSeq 2000 sequencing; Effect of MHC II expression in pro-B cells: Pre-B cells and follicular B cells from wild-type and MHC II conditional mice
design: Effect of MHC II expression in pro-B cells: Pre-B cells and follicular B cells from wild-type and MHC II conditional mice
library_construction_protocol: Mice carrying a conditional H2-Ab1 allele (H2-Ab1c) (Hashimoto, K., Joshi, S. K. & Koni, P. A. A conditional null allele of the major histocompatibility IA-beta chain gene. Genesis 32, 152-153 (2002)) were crossed to mice expressing Cre in DCs under the Itgax (CD11c-Cre driver; Itgaxcre) ( Caton, M. L., Smith-Raska, M. R. & Reizis, B. Notch-RBP-J signaling controls the homeostasis of CD8- dendritic cells in the spleen. J. Exp. Med. 204, 1653-1664 (2007)). This Cre transgenic mouse strain will delete the gene encoding the H2-Ab chain in all dendritic cells, as well as in a proportion of other hematopoietic cells. MHC II expression is developmentally regulated and begins in the bone marrow. Cells that have not yet expressed MHC II and those that have started expressing were compared from both wild-type and ItgaxcreH2-Ab1c MHC II conditional mice. Bone marrow pre-B cells (Hardy's Fraction D) and splenic follicular B cells were purified by flow cytometry according to MHC II protein expression from wild-type and ItgaxcreH2-Ab1c MHC II conditional mice and used for the isolation of RNA. RNA was then extracted using the RNeasy Mini QIAcube or Micro kits (Qiagen)  Illumina, RNA with PolyA selection (performed by GeneWiz)
attributes.tag: Experimental Factor: cell type
attributes.value: Pre-B cell
attributes.tag: Experimental Factor: genotype
attributes.value: ItgaxcreH2-Ab1c
attributes.tag: Experimental Factor: phenotype
attributes.value: MHC II-positive