Genetically Targeted Immunization and Metabolite Uptake in Mice

Use of B1–8i+/^– and B1–8i+/− J_κ^–/^– genetically targeted BALB/cJ mice was as described^{35 (link),36 (link)}. Mice were maintained under specific-pathogen-free conditions and all animal experiments were approved by the University of Pittsburgh Institutional Animal Care and Use Committee. 6- to 12- week-old mice were immunized i.p. with 50 μg of NP-CGG precipitated in alum to induce GCBCs or NP-Ficoll (Biosearch Technologies) in PBS to induce in vivo activated B cells. Mice were i.p. injected once a day at d9 and d13 post NP-CGG immunization with 22 mg/kg bodyweight Etomoxir (Cayman Chemicals) in 5% DMSO in PBS and 11 mg/kg Thioridazine (Cayman Chemicals) in 0.9% NaCl solution. For in vivo metabolite uptake experiments mice were injected intravenously with 0.171 mg (500 nmol) 2-NBDG (in PBS) or 50 μg BODIPYTM FL C16 (in 2.5% DMSO in PBS) and sacrificed 35 or 60 min later, respectively.