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. Author manuscript; available in PMC: 2013 May 1.
Published in final edited form as: Appl Microbiol Biotechnol. 2012 Jan 25;94(4):1041–1049. doi: 10.1007/s00253-012-3893-7

Fig. 3.

Fig. 3

Inhibitory effect of R9LF-1 on LF. (a) Protective effect of R9LF-1 against LF toxicity in J774A.1 cells. Cells were pre-incubated for 30 min with R9LF-1 at concentrations of 0, 30, and 50 μM before the treatment with lethal toxin composed of 500 ng LF and 500 ng PA per milliliter of medium. Cell viability was monitored by CCK-8. (b) Protective effect of R9LF-1 on MAPKK against LF in cell lysate. J774A.1 cell lysate was incubated with R9LF-1 at concentrations of 0, 10, and 30 μM in the presence of LF at 10 nM for 30 min at 37 C. MAPKK cleavage by LF was probed by monoclonal antibody against N-terminal MAPKK. (c) Protective effect of R9LF-1 on MAPKK against LF in living cells. J774A.1 cells were treated with R9LF-1 or medium 30 min prior to the addition of LT (500ng PA and LF per milliliter of medium). After 90 min incubation, MAPKK cleavage by LF was assessed by western blot.