Overexpression of the cholesterol-binding protein MLN64 induces liver damage in the mouse

Figure 4 Effect of recombinant MLN64 adenovirus infection on hepatic expressions of various apoptosis-regulating genes in mice. A: For caspase-3 analysis, liver cytosolic fractions were prepared from Ad.MLN64-infected mice, and a liver cytosolic fraction from a CDCA-fed mouse was used as a positive control (C+). For caspase-12 analysis, liver homogenates (TH) from two mice in each group were fractionated into total membranes (TM) and cytosol (CYT). Total liver extracts were prepared for the analysis of p53, p21, and Bax expressions. Proteins were size fractionated by SDS-PAGE, immunoblotted with anti-caspase-3, anti-caspase-12, anti-p53, anti-p21, and anti-Bax antibodies, and subjected to densitometric analysis. The protein expression data are shown after normalization to the ε-COP or albumin signal. ^aP < 0.05; B: Relative quantitative RT-PCR in the presence of [α-³²P]dCTP was used to analyze the bax and mdm2 RNA expressions. A 18S primer control was included in each sample. The products were resolved on a 1.5% agarose gel and transferred to a nylon filter, and the radiolabeled signals were quantified. Results are representative of three independent experiments. ^aP < 0.05.