TRAF6 is degraded by the autophagy lysosome pathway. (A) The indicated cell lines were cultured with 10nM bortezomib and cotreated with 5mM 3-MA for 24 hours. TRAF6 and GAPDH protein was determined by immunoblot analysis. (B) In the absence of proteasome inhibition, TF-1 cells were treated with 5mM 3-MA for 24 hours, and TRAF6 protein expression was measured by immunoblot analysis. (C) cDNA from TF-1 cells treated with 3-MA (5mM) or 3-MA and bortezomib (10nM) was evaluated for TRAF6 mRNA expression. Data are relative to GAPDH and normalized to cells treated with DMSO. (D) Cell viability of THP1 and TF-1 cells was determined after treatment with DMSO, 10nM bortezomib, or bortezomib and 5mM 3-MA for 24 hours. *P < .05.