Figure 4.

Human serum amyloid P (hSAP) pre-treatment decreases the carbon tetrachloride (CCl₄)-induced apoptosis of hepatocytes in vitro. NCTC 1469 cells were pre-treated with 30 μg/ml hSAP or the vehicle for 4 h and were then incubated with CCl₄ (2.5 mM) for 4 h. (A) Cells were trypsinized and stained with Annexin V and 7-AAD followed by analysis with flow cytometry. Early apoptotic cells (Annexin V positive and 7-AAD negative) are in the right lower quadrant. Late apoptotic or necrotic cells are in the right upper quadrant. The ratio of early apoptotic cells to total cells ×100% is shown. (B) The mRNA expression of B cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 associated X protein (Bax) from cells was detected using quantitative PCR. β-actin was used as a loading control. (C) The protein level of Bcl-2, Bax and cleaved caspase-3 from cells was detected by western blot analysis. (D) The expression levels of Bcl-2, Bax and cleaved caspase-3 were normalized to β-actin. ^**P<0.01. The results shown represent 3 independent experiments.