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. 2011 Nov 8;287(1):286–298. doi: 10.1074/jbc.M111.294355

FIGURE 6.

FIGURE 6.

DMBT1 induction through IL-27 is mediated by p38 and STAT3 and negatively regulated by STAT1 and STAT6. A, DMBT1 mRNA is induced by IL-27 and can be inhibited by treatment with p38 inhibitor SB203580 and STAT3 inhibitor S3I-201. DMBT1 expression in unstimulated control cells was arbitrarily set to 1.0. *, p < 0.01 versus control; **, p < 0.05 versus IL-27 only. PD, PD98059; SB, SB203580; Wo, wortmannin; S3I, S3I-201. B, DMBT1 protein is induced by IL-27 after 48 h. This effect is abolished by the p38 inhibitor SB203580 and the STAT3 inhibitor S3I-201. C, DMBT1 mRNA is induced 10-fold by IL-27 after 6 h in control siRNA-transfected cells. Silencing of STAT3 inhibited this effect, and silencing of STAT6 and even stronger silencing of STAT1 increased expression of DMBT1 mRNA induced by IL-27. D, DMBT1 protein is induced by IL-27 in control siRNA-transfected cells. STAT3 silencing abolished and STAT1 and STAT6 silencing increased expression of DMBT1 protein mediated by IL-27. Representative STAT1, STAT3, and STAT6 blots show silencing of the respective genes by the respective specific siRNA (C, control; S1, STAT1; S3, STAT3; S6, STAT6). E, Western blots analyzing nuclear extracts from IL-27-treated cells show a transfer of STAT3 protein into the nucleus. The nuclear protein p84 was used as loading control. F, ChIP analysis with an antibody against STAT3 demonstrates increased STAT3 binding to the DMBT1 promoter region (between bp −266 and −144 relative to the translation start site) in IL-27-stimulated cells. The negative control antibody (normal rabbit IgG antibody) did not produce any significant signal. *, p < 0.05 versus unstimulated.