FIGURE 2.

Knockdown of SREBP-1 reduces LPIN1 expression in Huh7 cells. Huh7 cells were transfected with 25 nm control (cont), SREBP-1 (S-1), or SREBP-2 (S-2) siRNA under LPDS conditions. A, C, and D, total RNA was extracted for analysis by real time RT-PCR 24 h after transfection. All values are expressed as means ± S.E. (n = 3). Target mRNA levels were normalized to cyclophilin A mRNA and are expressed relative to the values for cells transfected with control siRNA (cont), set at 1. The statistical significance of SREBP siRNA versus control siRNA is indicated as follows: * (p < 0.05) or ** (p < 0.01). N.S., not significant. B, 15 h after transfection, N-acetyl-Leu-Leu-norleucinal was added to a final concentration of 25 μg/ml, and the cells were harvested 1 h later. Aliquots (10 μg of protein/lane) were subjected to SDS-PAGE, and immunoblots were probed with anti-SREBP-1, anti-SREBP-2, or anti-lamin B2 antibody. The experiments were repeated three times. E, 48 h after transfection, cell extracts were prepared, and aliquots (50 μg of protein/lane) were subjected to SDS-PAGE. Immunoblots were probed with anti-lipin-1 antibody or anti-lamin B2 antibody. The experiments were repeated three times.