Effects of MTs siRNA on the expression of MTs and adipocyte-related genes and proteins.

<p>3T3-L1 preadipocytes were treated with 50 nM control RNA (open circle) or MTs siRNA (closed circle) 24 h before the addition of DIM (day -1). Total RNA was isolated on day 0, 1, 4, and 8, and the expressions of (A) <i>mt1</i>, (B) <i>mt2</i>, (C) <i>pparγ</i>, (D) <i>fabp4</i>, (E) <i>cebpβ</i>, and (F) <i>cebpδ</i>, were determined using real-time PCR. All mRNA levels were normalized to the expression level of the <i>36B4</i> gene and are shown as fold induction from the control RNA-treated cells on day 0. Data are expressed as the mean ± SD (<i>n</i> = 3). ^**<i>P</i> < 0.01, compared with control RNA-treated cells at the same time point. (G) Cell lysates from the control RNA-treated cells (control) and the MTs siRNA-treated cells (MTs siRNA) were obtained at the indicated points. The protein expression of PPARγ, FABP4, C/EBPβ, C/EBPδ, and FAS (with 36B4 as a loading control) during the adipogenic differentiation of 3T3-L1 cells was determined using immunoblotting.