Figure 3

E. adenophorum-induced apoptosis is mediated through the activation of caspase-9 and caspase-3.

(A)The protein levels of procaspases-3, -8 and -9 and the cleaved forms of these proteins. The expression of apoptosis-related proteins, including caspases-3, -9 and -8, is shown using β-actin as a control, as detected through western blot analysis. Densitometric quantification of the ratio of procaspases-3, -8 and -9 and the cleaved forms of them to β-actin. Data are normalized to control. (B) E. adenophorum administration induced apoptosome formation. Protein extracts from splenocytes were collected and used in immunoprecipitation assays against Apaf-1. The levels of caspase-9 and Cytc were detected through western blotting to indicate the formation of the apoptosome complex. (C) Relative mRNA levels of caspases-3, -8 and -9. The Saanen goats were treated with different doses of E. adenophorum for 3 months and mRNA was then extracted from splenocytes and used for qRT-PCR analysis. E. adenophoruminduced the activation of caspases-3 and -9. (D) Caspase activity in E. adenophorum-treated splenocytes. The BCA assay was used to verify equalamounts of protein and the enzymatic activities of caspases-8, -9 and -3 were measured using the colorimetric assay kit.(E) The concentration of caspases-3, -8 and -9 in splenocytes was measured through ELISA. The data are presented as the means ± SD of three independent experiments. *p < 0.05 and **p < 0.01, compared with the control group.