FIG 8.

WNT5A-Ca²⁺/CAMKII signaling axis regulates dectin-1-induced PIAS-1 and SOCS-1 expression. (A) Immunoblotting analysis for pCAMKII, Pyk2, and pTAK1 in macrophages treated with curdlan for the indicated time intervals. (B) Total cell lysates from peritoneal macrophages transfected with Wnt5a siRNA and treated with curdlan were analyzed for pCAMKII, pPyk2, and pTAK1 levels. (C to E) RAW 264.7 macrophages transfected with the TAK1 KN construct (C) or Tak1 siRNA (D and E) were treated with curdlan for 12 h. Total cell lysates were analyzed for PIAS-1 and SOCS-1 (C and D) and TAK1 (E) by immunoblotting. Mouse peritoneal macrophages from C3H/HeJ mice were pretreated with the indicated pharmacological inhibitors followed by 12 h (F) or 60 min (G) of treatment with curdlan to estimate the expression of PIAS-1 and SOCS-1 from whole-cell lysate (F) or nuclear translocation of NF-κB from nuclear and cytosolic fractions (G). (H) Analysis of curdlan-induced PIAS-1 and SOCS-1 expression in macrophages pretreated with NF-κB inhibitor, BAY 11-7082. All blots are representative of 3 independent experiments. NT, nontargeting; KN, kinase negative.