Figure 3.

PE6 induces the activation of the canonical NFκB signaling pathway. (A) Western blots showing the expression level of NFκB1 (P50) subunit of NFκB in PE6 treated RAW264.7 macrophage cells. RAW264.7 cells were treated with PE6 (1 and 2 µg/ml) for 24 h. (B) Densitometric quantitation of the P50 subunit of canonical NFκB. P50 protein levels were expressed relative to GAPDH [%]. (C) Immunofluorescence microscopic pictures showing nuclear translocation of P50 subunit. (D) Western blots showing the protein levels of the phosphorylated pP65 (RelA) subunit of NFκB in PE6 treated RAW264.7 macrophage cells. (E) Densitometric quantitation of pP65 subunit. The pP65 protein levels were expressed relative to GAPDH [%]. (F) Immunofluorescence microscopic pictures showing nuclear localization of phosphorylated P65. DAPI was used to mark the nucleus. Untreated and HI PE6 treated cells were used as negative controls. LPS (500 ng and 1 µg/ml) treatment was used as a positive control. GAPDH was used as a loading control. A488 linked secondary antibody was used for signal detection. Scale bars indicate 10 µm. Data were analyzed by one-way ANOVA with Tukey’s multiple comparison post-test. Data are representative of three independent experiments. *P < 0.05 and **P < 0.01 vs. controls.