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. 2003 Mar;108(3):391–400. doi: 10.1046/j.1365-2567.2003.01573.x

Figure 7.

Figure 7

Regulatory effects of MS-5 fibroblasts or thymic FCM on the transcriptional activity of IL-12p40 and p35 genes and IL-12 protein production by DC. Each figure is composed of histograms showing the IL-12p35 or p40 copies number per 1000 HPRT, and of symbols (×) showing the level of IL-12p70 (pg/ml) or IL-12p40 proteins (ng/ml). Day 12 cultured splenic DC (1 × 106/ml) were cultured or not on MS-5 fibroblasts monolayers (a) or in the presence of 20% thymic FCM (b) for 20 hr and then stimulated with DS-1 for a further 9 hr. (c) Ex vivo purified splenic CD11c+ cells (1 × 106/ml) were cultured on MS-5 fibroblast monolayers for 20 hr and then stimulated for a further 9 hr with PSB-2. Messenger RNA were extracted, reverse transcribed and amplified by real-time PCR using primers specific for IL-12p40, p35 and HPRT. IL-12p40 and p35 mRNA levels were quantified and normalized against HPRT mRNA using an internal standard curve of HPRT. Simultaneous measurements of IL-12p70 and IL-12p40 proteins were performed on culture supernatants using ELISA.