Figure 2.

Silencing C5aR with C5aR siRNA in vivo. A: Gene silencing in kidney detected by real-time PCR. 50 μg of C5aR-siRNA were injected i.v. into CD1 mice as described in Materials and Methods. 48 hours after gene silencing, kidneys were clamped for I/R injury. 24 hours after reperfusion, kidney tissues were harvested, and total RNAs were extracted. Transcripts of C5aR and GAPDH were determined by quantitative real-time PCR. C5aR expression was compared among the unclamped mice, vector treated and siRNA treatment I/R mice (n = 6, per group). Relative quantity of C5aR mRNA was expressed as mean ± SEM. Statistical significance as compared with controls (**) was denoted at P < 0.05. B: Complement component expression in kidney detected by immunochemistry. Mice were induced I/R as described above (A). Kidney tissues were taken from the unclamped control mice (a, d, g), control vector treated I/R mice (b, e, h), or C5aR-siRNA treated I/R mice (c, f, i). Frozen sections were stained with antibodies against C5aR (a, b, c), C3 (d, e, f), or C9 (g, h, i).