Figure 4

WB analysis of Wnt/NF-κB interplay. A cross talk between TLR4 and Wnt signaling occurred as demonstrated by WB analysis at T0, T30′, T1h, and T2h using specific antibodies for cytoplasmic p(Ser473)-Akt, p(Ser9)-GSK3β, p(Ser33)-β-catenin and nuclear β-catenin (n-β-catenin), and NF-κB p65 (n NF-κB p65) in unstimulated cells; (A) samples treated with Wnt3a (20 ng/mL) (B) and LPS (5 μg/mL) (C) in SM and BM medium. GAPDH and lamin B were considered as housekeeping proteins for the analysis of cytoplasmic and nuclear antigens. The quantification of protein expression levels was performed using ImageJ processing software. Data were reported as ratio within target protein and relative housekeeping protein expression. Statistical significance was calculated by Student’s t-test by comparing to control: p value ≤0.05*, p value ≤0.01**.