Fig. 5

IL-1β induces Saa3 expression in alveolar epithelial cells in an NF-κB dependent manner. (A) Intravenous administration of IL-1β induced Saa3 expression in the lung. Detection of Saa3 mRNA by qPCR 6 and 24 h after IL-1β administration (20 ng in 100 ul saline). (B) The effect of IL-1β on the expression of Saa3 in different cell lines. Detection of Saa3 mRNA by qPCR 6 and 24 h after IL-1β (10 ng/ml) stimulation on MLE-12, RAW264.7 and MS1 cells. (C) Silencing of Il1r1 blocked IL-1β-induced Saa3 expression in vivo. Lenti-shNC or lenti-shIl1r1 viruses were given intratracheally to the lung of WT mice. After 7 days, mice were intravenously given saline or IL-1β (20 ng in 100 ul saline). After 6 h, Saa3 and Il1r1 expression was detected by qPCR. (D) Silencing of Il1r1 blocked IL-1β-induced Saa3 expression in MLE-12 cells. Forty-eight hours after siRNA transfection, MLE-12 cells were stimulated with IL-1β (10 ng/ml) for 6 h. Saa3 mRNA level was detected by qPCR. (E) The effect of NF-κB, ERK, JNK and p38 inhibitors on IL-1β-induced Saa3 expression in vivo. One hour after intravenous administration of indicated inhibitors, IL-1β (20 ng in 100 ul saline) were i.v. administrated into the mice. Six hours later, lung Saa3 expression was detected by qPCR. (F) The effect of NF-κB, ERK, JNK and p38 inhibitors on IL-1β-induced Saa3 expression in MLE-12 cells. MLE-12 cells were pre-treated with indicated inhibitors for 1 h, then stimulated with IL-1β (10 ng/ml) for 6 h. Saa3 expression was detected by qPCR. (G) Silencing of p65 blocked IL-1β-induced Saa3 expression in MLE-12 cells. Forty-eight hours after siRNA transfection, MLE-12 cells were stimulated with IL-1β (10 ng/ml) for 6 h. Saa3 mRNA level was detected by qPCR. Data are displayed as the mean ± SEM; (A, E and F) one-way ANOVA with Turkey’s correction. (B) one-way ANOVA with Dunnett’s correction. (C, D and G) two-way ANOVA with Turkey’s correction. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant