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. 2016 Apr 20;6:24575. doi: 10.1038/srep24575

Figure 5. Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.

Figure 5

(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.