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. 2016 Feb 10;7(18):25162–25179. doi: 10.18632/oncotarget.7301

Figure 5. DHA inhibits TPA-induced PKCδ activation, STAT3 DNA binding activity, β-catenin, and STAT3α expression.

Figure 5

MCF-7 cells were pretreated with 0, 25, or 100 μM DHA for 24 h followed by incubation with 100 ng/ml of TPA for another 30 min. PKCδ in the plasma membrane and cytosol (A) and STAT3α (B) and GSK3β (C) phosphorylation were determined. (D) Total cellular β-catenin and STAT3α and nuclear β-catenin protein levels were determined in cells pretreated with various concentrations of DHA for 24 h followed by incubation with TPA for another 24 h and 4 h, respectively. (E) Cells were pretreated with 100 μM DHA for 24 h, 5 μM WP for 4 h, or 5 μM rottlerin (Rot) for 1 h, and then were incubated with TPA for another 6 h. Nuclear extracts (10 μg) were prepared for STAT3 nuclear protein DNA binding activity assay. To confirm the specificity of the nucleotide, 50-fold cold probe and mutant (Mu) were included in the EMSA. One representative experiment out of three independent experiments is shown. **p < 0.01.