a. Q-RT-PCR analysis of A20 mRNA levels of YFP+ LV-siA20-DC and LV-siGFP-DCs after stimulation with LPS for 12 hr. Experiments were repeated twice with similar results. *P < 0.01, vs. siGFP-DC. Schematic diagram of the recombinant lentiviral vector LV-siA20 (top panel) is presented. H1, H1 RNA promoter; cPPT, central polypurine tract sequence; IRES, internal ribosome entry site; wPRE, Woodchuck post-transcriptional regulatory element sequence; and siA20, A20 small hairpin interfering RNA.
b. Surface expression of molecules on CD11c+/YFP+ gated BM-DC 48 h after viral transfection using FACS analysis. Experiments were repeated three times with similar results.
c. ELISA measurement of the levels of cytokines secreted by transfected DCs (5×105 cells/ml) in response to stimulation with LPS (100 ng/ml) for 16h from one representative experiment of three. *P < 0.01, vs. siA20-DC.