Partially purified βh/c triggers MAPK activation and inhibits IL-12 and NOS2 expression in macrophages.

<p><b>A</b>. βh/c extract was prepared from wt GBS (10/84; βh/c^e) and a mock extract from 10/84Δ<i>cylE</i> bacteria (ΔcylE^e), hemolytic activity measured by sheep red blood cell (RBC) lysis assay. <b>B</b>. Macrophages were stimulated with heat-killed 10/84Δ<i>cylE</i> bacteria (h10/84Δ, MOI: 20) in the presence of βh/c^e or ΔcylE^e at a dilution of 1∶200, and protein extracts prepared at the indicated time points. JNK and p38 activity were measured by IP kinase assay, as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002812#ppat-1002812-g002" target="_blank">Fig. 2</a>. IB for p38 was used as loading control. Representative data from at least 2 independent experiments is shown. <b>C</b>. IL-12p40 was measured in cell culture supernatants by ELISA at 8 and 24 h after stimulation of macrophages with h10/84Δ in the presence of βh/c^e or ΔcylE^e. Data is represented as mean ± sem of 4 replicates, representative data from at least 3 independent experiments is shown. <b>D</b>. Protein extracts were prepared from macrophages 8 h after stimulation with h10/84Δ in the presence of βh/c^e or ΔcylE^e and NOS2 expression measured by IB, ERK expression was used as a loading control. Representative data from at least 3 independent experiments is shown.