4C-seq analyses of the 3'Igh locus during B cell development

We report the application of 4C-seq for analysis of the 3' end of he Igh locus during B cell development. We generated genome-wide 4C maps for BM derived Rag2-/- pro-B cells, resting splenic B cells, LPS+IL4 activated B cells and ConA stimulated splenic T cells using two viewpoints, Em and hs4 located in 3'Ea. We find that long range interactions are present only on B lineage cells reflecting enhancer activation status. Overall design: Chromatin templates were isolated from Rag2-deficient B cell progenitors expanded ex vivo in IL7, column purified resting splenic B cells, B cells activated with LPS+IL4 for 40 hours and splenic T cells that were ConA activated. These samples were were then submitted to 4C-seq analysis, libraries prepared and submitted to NGS studies. Biological replicates were prepared.