B1-8hi DEC205+/+ GC B cells, anti-DEC-CS, G1 phase replicate 1
To generate replication timing data, cell suspensions from draining lymph nodes were stained for surface markers and then fixed and permeabilized as described above. Samples were then incubated in 100 µl of 20 µg/µl RNAse A (Life Technologies) for 30 minutes at 37°C and subsequently stained for DNA content with 7-AAD. B1-8hi DEC205+/+ GC B cells were identified as B220+CD38- FAS+CD45.1+CD45.2- and sorted based on G1 or S phase DNA content. Genomic DNA from sorted cells was isolated and libraries derived from these samples were sequenced on a HiSeq 2500 (Illumina)