Fig. 1.
In vitro stimulation of naïve cells isolated from TCR transgenic mice (OT-1/Rag −/−). The CD8+ T cells obtained from naïve TCR transgenic mice (OT-1/Rag −/−) mice are stimulated in vitro with latex microspheres on which major histocompatibility complex (MHC) Class I (H-2Kb) dimers bearing 10 nM of cognate peptide (SIINFEKL) (Ag) are immobilized, along with 1 μg/ml of recombinant murine B7.1 (co-stimulation) and 2 ng/ml of rmIL-12 (cytokine). The cells are harvested at the indicated time points and evaluated for activation, clonal expansion, survival, metabolic status and differentiation by standard methodologies including flow cytometry, western blotting, PCR and metabolic flux analysis. UCP2 is inhibited (UCP2 siRNA or Genepin) after antigen stimulation.