HEL treated samples were washed after 5 days, replaced with new media with no IL-7, but 1ug/mL HEL. Control samples were treated the same except no HEL in the media. HEL Mlo samples were sorted for IgMa low, IgDneg. HEL 48h samples were sorted for B220+ IgMahi, GFP+ cells after 48h in HEL. BM 48h Ctrl samples were sorted for B220+ IgMahi GFPneg after 48h in culture with no HEL.
Growth protocol
5 day culture in RPMI + 10% FBS, Pen/Strep, L-glut, 2-ME, 16ng/mL IL-7
Extracted molecule
total RNA
Extraction protocol
Qiagen RNeasy protocol
Label
biotin
Label protocol
Ambion cDNA synthesis with T7 oligo dT primer (Array Script). Labeling with biotin-11-CTP and biotin-16-UTP.
