1C). Hence, STAT3-induced β-catenin inhibits BMDCs maturation and function. To elucidate the regulatory role of STAT3 and β-catenin on DC function, we disrupted STAT3 signaling in BMDCs by using a small interfering RNA (siSTAT3). This resulted in diminished CoPP-/rIL-10-mediated β-catenin expression (Fig. 2A; 0.2-0.4 AU), compared with nonspecific (NS) siRNA-transfected

cells (2.5-2.8 AU). In addition, SnPP (HO-1 inhibitor)-treated cells showed decreased β-catenin levels (0.2-0.6 AU). Interestingly, specific NVP-BEZ235 purchase knockdown of STAT3 in CoPP-/rIL-10-treated BMDCs promoted PTEN activation (Fig. 2A; 2.2-2.4 AU) but inhibited Akt phosphorylation (0.2-0.5 AU), as compared with NS siRNA-transfected cells (0.2-0.3 AU and 2.3-2.5 AU, respectively). Furthermore, disruption of STAT3 reversed CoPP or rIL-10-mediated inhibition of LPS-triggered DC maturation, evidenced by increased CD40, CD80, and CD86 expression (Fig. 2B). Consistent

with flow cytometry data, the production of IL-12p40, TNF-α, and IL-6 was elevated after blockade of STAT3 in CoPP- or rIL-10-treated, but not in NS siRNA-treated BMDCs (Fig. 2C). Thus, STAT3 knockdown inhibits β-catenin signaling and triggers PTEN/PI3K and DC maturation, suggesting that β-catenin regulates DC function in a STAT3-dependent manner. To further dissect putative selleckchem mechanisms by which β-catenin may regulate DC function, we disrupted β-catenin signaling in BMDCs by using a small interfering RNA (siβ-cat). As shown in Fig. 3A, LPS-stimulated BMDCs readily induced PTEN (2.3-2.5 AU) and TLR4 (2.6-2.8 AU). Interestingly, disruption of β-catenin in CoPP or rIL-10 pretreated BMDCs led to enhanced expression of PTEN and TLR4 (2.2-2.4 AU and 1.9-2.1 AU, respectively) compared

to nonspecific siRNA (siNS)-treated controls (0.3-0.7 AU and 0.2-0.4 AU, respectively). Furthermore, knockdown of β-catenin in CoPP or rIL-10 pretreated BMDCs increased the phosphorylation of IRF3 and IκBα (Fig. 3A; 1.5-1.7 AU and 1.6-1.8 AU, respectively). Similar findings were recorded in LPS-stimulated BMDC without adjunctive CoPP/rIL-10 (Supporting Fig. 3). As PTEN/PI3K signaling regulates TLR4 activation in DCs,24 we used the PTEN phosphate release assay, in which β-catenin knockdown almost was found to increase PTEN activity (Fig. 3B) in CoPP- or rIL-10-treated LPS-stimulated DCs. These results were consistent with increased expression of CCR2, CCR5, and CXCR3 in siβ-cat-treated DCs, compared with those without β-catenin-silenced cells (Fig. 3C). Thus, disruption of β-catenin activates PTEN/TLR4 signaling in DCs. Next, we investigated whether disruption β-catenin signaling may affect local inflammatory responses in a mouse liver IRI model. The hepatocellular damage at 6 hours of reperfusion following 90 minutes of partial liver warm ischemia was evaluated by Suzuki’s histological grading (Fig. 4A/B).