Figure 4.

In vitro treatment of LPS-activated rodent microglia with Red extract reduces the production of pro-inflammatory mediators. Rodent microglia cells were grown in culture, overnight exposed to medium alone, Yellow or Red extracts, and then challenged with medium alone or 1 μg/ml LPS for 6 h (see “Materials and Methods” section for details). Cells were then collected and the expression of pro-inflammatory cytokine genes was evaluated by Real time RT-PCR (A). Each transcript was normalized to corresponding control value (V, vehicle). Values shown are mean ± SEM from two independent experiments performed in triplicate. (B) Evaluation of iNOS protein levels by Western blotting and relative quantification. Filter is from one representative experiment out of three (see Supplementary Figure S2 for the specificity of antibodies). Densitometric analysis of bands have been performed with ImageJ and values normalized to α-tubulin. Histograms show the mean ± SEM values from three independent experiments performed in triplicate. *p < 0.05, **p < 0.01, and ***p < 0.001 with respect to cultures exposed to Red extract, one-way ANOVA followed by Bonferroni post hoc tests.