title: Illumina HiSeq 4000 paired end sequencing; RNAseq of follicular lymphoma patient samples treated with anti-IgM, DC-SIGN or an isotype control
design: RNAseq of follicular lymphoma patient samples treated with anti-IgM, DC-SIGN or an isotype control
library_construction_protocol: Primary FL cells were used following approval from the Southampton and South West Hampshire Research Ethics Committee and Rochester, and with informed consent, in accordance with the Declaration of Helsinki. Cells were recovered from cryopreservation as previously described (6). Tumor IGHV gene sequences were identified as described (6). Where cells were incubated with DC-SIGN-Fc or DC-SIGN-HA (both R&D Systems) media was additionally supplemented with CaCl2 to a final Ca2+ concentration of 2 mM, unless otherwise stated. DC-SIGN-Fc/HA were used at 20 µg/ml and goat anti-human F(ab')2 anti-IgM (anti-IgM) and goat F(ab')2 IgG control (both Southern Biotech) were used at 10 µg/ml, unless otherwise stated.  FL samples were treated with 20 μg/ml of goat F(ab')2 anti-IgM, 20 μg/ml of DC-SIGN-Fc or left untreated for 4 hours at 37°C.  Total RNA was extracted using an RNeasy mini kit (Qiagen) PolyA