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. 2003 Aug 15;112(4):598–607. doi: 10.1172/JCI17129

Figure 3.

Figure 3

Regulation of IL-6 production by IL-1β and IFN-γ in vitro and in vivo. Growth-arrested HPMC were (a) incubated with medium alone or IL-1β (100 pg/ml) in the presence or absence of IFN-γ (100 U/ml). At specific time points, IL-6 levels were quantified using ELISA (*P < 0.05, a significant increase versus the additive value of IL-1β plus IFN-γ alone). (b) Northern blot analysis of total RNA isolated from HPMC incubated for 3 hours with medium and IL-1β (100 pg/ml) in the presence or absence of IFN-γ (100 U/ml). Representative results for three separate experiments are shown. (c) HPMC were stimulated with medium alone (lane 1), HYPER-IL-6 (500 pg/ml, lane 2), IFN-γ (100 U/ml, lane 3), IL-1β (100 pg/ml, lane 4), or a combination of IL-1β and IFN-γ (lane 5). At the indicated intervals, nuclear extracts were prepared and NF-κB activation was monitored by EMSA. (d) Composition of the NF-κB complex was determined by supershift using antibodies against p50 (lane 2) and p65 (lane 3). No antibodies were included in lane 1. Results are representative of three separate experiments performed with HPMC from different donors. (e) Wild-type mice were intraperitoneally administered with PBS, IL-1β (100 ng per mouse), IFN-γ (0.5 U per mouse), or a combination of IL-1β and IFN-γ at these doses. After 1 hour, mice were sacrificed and IL-6 was quantified (*P < 0.05 versus PBS alone; **P < 0.05, a significant increase versus the additive value of IL-1β plus IFN-γ alone).