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. 2018 Sep 10;16(9):323. doi: 10.3390/md16090323

Figure 6.

Figure 6

Effects of fucoxanthin (Fx) on signal protein-1 (SP-1) activity in TGF-β1-induced NPDFs. (A) Cells were pretreated with Fx (5–30 µM) for 1 h before stimulation with TGF-β1 (1 ng/mL) for 30 min. Nuclear extracts were prepared and analyzed for SP-1 activity via western blotting. (B) Cells were incubated with 20 μM LY294002 for 1 h followed by TGF-β1 treatment for 30 min, and nuclear extracts were prepared and analyzed for SP-1 activity via western blotting. (C) After pretreatment with 20 μM LY294002 for 1 h, the nuclear proteins were isolated after a 30 min interval and Smad 2 and Smad 3 nuclear translocation was determined via western blot analysis using antibodies specific to the phosphorylated forms of Smad 2 and 3. (D) The NPDFs were transfected with either the control siRNA (80 nM) or the Smad 2/3 siRNA (80 nM) for 24 h, followed by stimulation with TGF-β1 (1 ng/mL) for 30 min. Untreated cells were used as a control (CON).