Effects of fucoxanthin (Fx) on TGF-β1-induced phosphorylation of ERK-1/2, SAPK/JNK, p38 MAP kinase, and Akt in NPDFs. The NPDFs were treated with the indicated concentrations of Fx for 2 h, followed by stimulation with TGF-β1 (1 ng/mL) for 30 min. (A) Cell extracts were prepared and subjected to western blotting with antibodies specific for the phosphorylated forms of ERK, JNK, p38, and Akt. The results are representative of three independent experiments. (B) After treatment with TGF-β1 for 24 h in the presence or absence of LY294002 (LY) (20 μM), the cell extracts were isolated and western blot analysis of α-SMA and Col-1 proteins was performed. Untreated cells were used as a control (CON). ERK-1/2, extracellilar signal-regulated kinase-1/2; SAPK/JNK, Stress-activated protein kinase/c-Jun NH2-terminal kinase; p38 MAP kinase, p38 mitogen-activated protein kinase.