Fig. 4.

Effect of DHA or insulin on the pERK1/2/ERK1/2, pAkt/Akt, and pS6/S6 ratios and p-SREBP-1, m-SREBP-1, and FASN expression in MCF-7 cells. Cells were pretreated with BSA or 60 μM BSA-bound DHA for 48 h in DMEM containing 5% FBS, then the same medium alone or 1 μg/ml of insulin was added. Western blots were then used to measure the pERK1/2/ERK1/2, pAkt/Akt, and pS6/S6 ratios after 1 h and p-SREBP-1, m-SREBP-1, and FASN expression after 24 h (a). Total ERK1/2, Akt, or S6 was used as the loading control for pERK1/2 (b), pAkt (c), or pS6 (d), respectively, while GAPDH was used as the loading control for p-SREBP-1 (e), m-SREBP-1 (f), and FASN (g).The levels are expressed as a fold value compared to the BSA-treated control with no insulin stimulation. Two-way ANOVA followed by the Bonferroni posttest was used to compare DHA and insulin effects. The data are presented as the mean ± S.E.M for 5–6 independent experiments