Fig. 2

Application of anti-TLR for the analysis of molecular mechanisms of TLR. Expressed TLRs are transported from ER to plasma membrane or endolysosome (1 and 2). In endolysosome, nucleic acid-sensing TLRs are cleaved and the N-terminal fragment binds to the C-terminal fragment (3). After the modification, TLR recognize ligand and signaling pathway is activated (5). TLRs on cell surface recognize ligand and activates signaling pathway. Some of them are internalized by the recognition of ligand or spontaneous trafficking (4). For the analysis of these molecular behaviors of TLR, monoclonal anti-TLR is well exploited. Amount and expression pattern among cell types are analyzed by flowcytometry. Subcellular distribution is observed by immunofluorescence microscopy. Alternatively, flowcytometry following the immunostaining with or without detergent is able to discriminate the distribution between cell surface and intracellular particle. Proteolytic cleavage and binding of nucleic acid-sensing TLR are detected by immunoprecipitation and immunoblotting. In addition to these application for detection, several antibodies have inhibitory effect on the response of TLRs. Words in black frames, blue frames, and orange frames are the name of organelle, the molecular behavior of TLR, and the application of anti-TLR, respectively