Figure 4. B. anthracis induces ATP release from infected macrophages in LF-, p38- and AKT-dependent manner.

(A) C57BL/6 macrophages were infected with WT (BA) or ΔLF B. anthracis. At the indicated times points, ATP in the culture supernatants was measured. This experiment was repeated at least 4 times and the results of one representative experiment done in triplicates are shown. Results are means ± SD, *p < 0.05, **p < 0.01 denote significant differences between the groups.

(B,C) Mouse macrophages were infected with WT (BA) or ΔLF B. anthracis in the absence or presence of KN-62 (10 μM). After 8 hr, IL-1β (B) and IL-6 (C) in culture supernatants were measured by ELISA. This experiment was repeated 3 times and the results of one representative experiment done in triplicates are shown. Results are means ± SD, **p < 0.01 denote significant differences between the groups.

(D) RAW264.7 cells transfected with MEK6ΔCR, myr-AKT or empty vector were left uninfected or infected with B. anthracis and 2 hr later ATP release to culture supernatants was measured. This experiment was repeated 3 times and the results of one representative experiment done in triplicates are shown.

(E) C57BL/6 macrophages were infected with WT B. anthracis in the absence or presence of 130 mM KCl and 8 hr later the amount of IL-1β in culture supernatants was determined by ELISA. This experiment was repeated 3 times and the results of one representative experiment done in duplicates are shown. Results are means ± SD, **p < 0.01 denote significant differences between the groups.

(F) Mouse macrophages were infected with WT B. anthracis in the absence or presence of KN-62 (10 μM) or KCl (130 mM) and at the indicated times cell lysates were prepared and analyzed by immunoblotting of 2 separate gels from the same experiment for MEK proteolysis. This experiment was repeated 3 times and the results of one representative experiment are shown.