GA affects the phenotype of MO-DCs in a gene-dependent manner. Aliquots of MO-DCs on day 6 of culture were differentially treated with GA (0.1 μM) and/or stimulation cocktail (see Methods section) as indicated for 48 h. (a) The expression levels of the markers indicated were assessed by flow cytometry. Upper panel: Marker expression was detected in unstimulated (-) and cocktail-stimulated (stim) MO-DCs left untreated (dark line) or treated with 0.1 μM GA (light grey). Shaded area: isotype control of MO-DCs left untreated (corresponding isotype controls of GA-treated MO-DCs were comparable). Graphs are representative of 4-5 independent experiments each. Lower panel: Relative expression intensities of DC surface marker expression are given as mean fluorescence intensities (MFIs), normalized to the MFI of unstimulated or stimulated MO-DCs left untreated. Data represent the means ± SEM of 4-5 independent experiments each. (b) Contents of IL-6 and IL-12p40 in the supernatants of harvested MO-DC populations were assayed by ELISA. Data represent means ± SEM of 10 independent experiments each. nd: not detectable. Statistical significance: (a) *versus untreated MO-DCs, (b) *versus unstimulated untreated MO-DCs, #GA-treated at stimulated versus unstimulated state. (a, b) *P < 0.05, ##,**P < 0.01, ***P < 0.001.