Figure 6

Fisetin promotes the expression of β-catenin and its nuclear translocation in LPS-stimulated RAW 264.7 macrophages. RAW 264.7 macrophages (1 × 10⁵ cells/mL) were pretreated with fisetin 2 h prior to LPS treatment (500 ng/mL; 0.5 h). (A) Total proteins were isolated and equal amounts of proteins were resolved on SDS–polyacrylamide gels, transferred to PVDF membranes, and probed with antibodies against total GSK-3β, phosphor-GSK-3β (Ser9), and β-catenin. β-Actin was used as an internal control. (B) In a parallel experiment, nuclear proteins were isolated and western blotting was performed to verify the nuclear translocation of β-catenin. Nucleolin was used as an internal control. (C) Nuclear translocation of β-catenin was detected by immunofluorescence. Each value indicates the mean ± standard error median (SEM), and is representative of the results obtained from three independent experiments. Significant differences among the groups were determined using Student’s t-test (^###p < 0.001 vs. untreated cells) and one-way ANOVA with Bonferroni correction (***p < 0.001 and *p < 0.01 vs. LPS-treated cells). UT untreatment.