Figure 5. Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.
(A,B) HMDM were stimulated with 10 nM C5a and lysed after 30 min. For antagonism of C5aR, HMDM were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to 10 nM C5a stimulation after (A) 1 h or (B) 16 h post-treatment with antagonist. (C) Rat macrophages were stimulated with 0.1 μM C5a and cells are lysed after 30 min. For antagonism of C5aR, cells were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to C5a treatment after 1 h post-treatment with antagonist. Gene expression was measured by real-time PCR, normalized against 18S and converted to fold change relative to control. Error bars are means ± SEM of three independent experiments (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.