Activated Dectin-2 and Dectin-3 can trigger themselves for ubiquitination and degradation. (A and B) Hyphae-induced ubiquitination and degradation of Dectin-2 and Dectin-3 in mouse BMDMs. (C and D) α-Mannan­–induced ubiquitination of Dectin-2 and Dectin-3 in RAW264.7 cells overexpressing Dectin-2 or Dectin-3. (E) α-Mannan–induced K48 ubiquitination of Dectin-2 and Dectin-3 in RAW264.7 cells overexpressing Dectin-2 and Dectin-3. (F) Hyphae-induced K48 ubiquitination of Dectin-2 and Dectin-3 in RAW264.7 cells overexpressing mutant Dectin-2_K12A and Dectin-3_K9A. Immunoprecipitation was performed using anti–Dectin-2 or –Dectin-3 antibodies on solubilized protein extracts from BMDMs or RAW264.7 cells and subjected to immunoblotting with the indicated antibodies. (G and H) Hyphae-induced degradation levels in RAW264.7 cells expressing WT or mutant Dectin-2 (G) and Dectin-3 (H), which were pretreated with 10 µg/ml cycloheximide for 15 min and then stimulated with hyphae (multiplicity of infection [MOI] =1) for the indicated times. The cell lysates were subjected to immunoblotting using the indicated antibodies. The data shown are representative of three independent and reproducible experiments. IP, immunoprecipitation; Ly, lysis; Ub, ubiquitination.