SMO activates NF-κB through PKCβ-CARMA1-MALT1-BCL10-TRAF6-NEMO axis. (A) Stably knocking down SMO or overexpressing SMO showed a decrease or increase in total PKC activity in HEK293T cells, respectively. Inhibition of SMO by cyclopamine-KAAD-KAAD in BJAB or silencing SMO in HBL1 also resulted in decrease of total PKC activity, whereas activating SMO by recombinant Shh in BJAB resulted in increase of total PKC activity. (B) Inhibition of SMO by cyclopamine-KAAD or silencing of SMO in DLBCL cell lines resulted in decrease of p-PKCβ-1 and p-PKCβ-2 and a decrease of p-CARMA1. In contrast, activating SMO by recombinant Shh resulted in increase of p-PKCβ-1 and p-PKCβ-2 followed by increased of p-CARMA1. (C) Activation of SMO with recombinant Shh increased the binding between SMO and CARMAL1/BCL10/MALT1/TRAF6 complex (central lane; +SMO). TCL, total cell lysates. (D) Transient overexpression of SMO resulted in increased TRAF6 and NEMO ubiquitination on K63 lysine indicating stabilization of these proteins and supporting NF-κB activation.