The relationship of bbtTICAM with other adaptors. (A) bbtTRAF2 attenuates the signal pathway induced by bbtTICAM-FL-WT in a dose-dependent manner. bbtTICAM-FL-ΔT2 is not affected by bbtTRAF2 as bbtTICAM-FL-WT. The pictures from western blot below the bar graph show expression of relevant proteins, including bbtTICAM, bbtTRAF2, and β-actin. (B) bbtTICAM-FL can be negatively regulated by bbtSARM in a dose-dependent manner. The pictures from western blot below the bar graph show expression of relevant proteins, including bbtTICAM, bbtSARM and β-actin. (C) Site-directed mutation of TRAF2-binding motif. PxQxS (TRAF2-binding site) is substituted for AxAxA. (D) bbtTICAM-FL-WT co-localizes with bbtTRAF2 in Hela cell line. (E) Co-IP results show that the truncated mutant bbtTICAM-N+TIR can interact directly with bbtTRAF2. When the TRAF2-binding motif was mutated, the interaction was attenuated. (F) Co-IP results show that the truncated mutant bbtTICAM-N+TIR can interact with bbtSARM directly. (G) Signal transduction and regulation of amphioxus MyD88-dependent and -independent pathways, setting up the foundation of vertebrate TLR signaling network.