Promotion of TLR7-MyD88-dependent Inflammation and Autoimmunity through Stem-loop Changes in Lnc-Atg16l1
In this study, we employed a combination of Endosome RNA-seq and iCLIP-seq technologies to identify long non-coding RNA associated with TLR7 and Myd88 in mouse HA-mTLR7 stable overexpression and wild type RAW264.7 cells. RNA-seqs of wild type and Lnc-Atg16l1 RAW264.7 cells were applied to function study. We performed icSHAPE-seq to investigate RNA secondary structure of Lnc-Atg16l1 in RAW264.7 cells with or without stumulatied by R837, as well as in spleen cells from BXSB.Yaa and BXSB.B6 mice. Overall design: RNAs from endosome in mouse HA-mTLR7 stable overexpression RAW264.7 cells  was analyzed using illumine Novaseq 6000. RNAs from RAW264.7 cells with or without R837 stimulating was analyzed using illumine Novaseq 6000. TLR7 activated iCLIP were performed using anti-HA antibody in RAW264.7 cells stably expressing HA-mTLR7. TLR7 noactivated iCLIP were performed using anti-TLR7 antibody in RAW264.7 cells. Two iCLIP of Myd88 were performed using anti-Myd88 antibody in RAW264.7 cells with or without R837 stimulating. icSHAPE experiments were performed through click reaction in RAW264.7 cells with or without R837 stimulating and spleen cells from BXSB.Yaa and BXSB.B6 mice. All RNAs of iCLIP and icSHAPE were analyzed using illumima Hiseq 2500.