Figure 5
Effects of PUFAs on IL-6-induced STAT3 signaling pathway. (A) Time-dependent effect of IL-6 on STAT3 levels in the nucleus and total cell lysates was measured by western blot analysis. The expression of B23 and α-tubulin was used as the internal control for the nuclear protein and cytoplasmic protein, respectively. (B) Immunoblot analysis of whole cell lysates from cells pretreated with 100 μM PUFAs for 24 h and followed by stimulation with IL-6 for 30 min. (C) Parallel experiments as in panel B, but detecting STAT3 phosphorylation in the nucleus. Data were quantified by densitometry and the value of the IL-6-treated group was set as 100%. The results are presented as the mean ± SEM of at least three independent experiments, **P < 0.01 compared to the IL-6-treated group. (D) Cells were treated with IL-6 in absence or presence of the protein tyrosine phosphatase (PTPase) inhibitor Na3VO4 for 1 h. The effects of DHA and EPA on the phospho-Tyr705-STAT3 levels in the nucleus were determined by immunoblot analysis. (E) Parallel experiments similar to those described in panel D, but observing the effect of LA and AA. The photograph depicts a representative gel from three independent experiments.