Figure 5. ESE-3b expression is increased in immunogenic moDC.

Monocytes were negatively selected from PBMC using magnetic beads. Immature moDC were generated with IL-4 and GM-CSF for 6 days. 15d-PGJ₂ (PGJ2 DC) and dexamethasone plus 1α, 25-dihydroxyvitamin were added to generate tolerogenic moDC, respectively (PGJ2 DC and Dex/VD3 DC). To generate immunogenic moDC, immature moDC were stimulated for 24 h with LPS, polyI:C and a cytokine cocktail containing TNF-α, IL-1β, IL-6 and PGE₂, respectively. (A) ESE3a and (B) ESE-3b mRNA levels were analyzed by quantitative real-time RT PCR using isoform specific primer/probes. GAPDH was used for normalization, immature moDC were used as reference. N = 4–6, *p<0.05. (C) Protein levels of ESE-3 in nuclear extracts were analyzed by Western blotting using a monoclonal antibody specific for human ESE-3. Histone H3 was included to ensure equal loading of the gel. One representative experiment out of three is shown.