DAF reduces interaction of C3a-C3aR and MAC formation in rat neurons subjected to hypoxic-ischemic conditions. Primary rat cortical neurons were exposed to NaCN for 1 hr then allowed to recover for 24 hours in normal medium with or without DAF. (a) Cell lysates were analyzed by immunoblot using anti-C3a and anti-β-actin antibodies. Data was quantified by densitometry (n = 5). (b) Interaction between neuronal C3a and C3aR was determined by immunofluorescent staining using anti-C3a (red) and anti-C3aR (green) antibodies (n = 3). Enlarged images are in the lower left boxes of merged images. (c) Cells were stained with mouse anti-rat MAC antibody (green), DAF resulted in a decrease in MAC formation (n = 3). Original magnification: ×400. Scale bar = 50 μm. (one-way ANOVA followed by Newman-Keuls test. *p value < 0.05, **p < 0.01).