Figure 2. Antimicrobial compounds (ROS and NO) enhance Perforin-2 mediated killing of S. typhimurium by PEM but have limited activity in the absence of Perforin-2.

(A–D) Wild-type S. typhimurium infection of PEMs isolated from either MPEG1 (Perforin-2) +/+ (A, C), or MPEG1 (Perforin-2) −/− mice (B, D). Non-filled symbols indicated MPEG1 (Perforin-2) +/+ PEMs; whereas filled symbols are MPEG1 (Perforin-2) −/− PEMs. Cells were incubated with NAC (blue line), NAME (green line), or mock (black line). To assess bacterial resistance mechanisms against these effectors, (E) SodC1 or (F) HmpA knockout S. typhimurium were used to infect MPEG1 (Perforin-2) −/− or +/+ PEMs. The above experiments were conducted with six biologic replicates and are representative of four independent experiments. Statistical analysis was performed utilizing multiple T-tests with correction for multiple comparisons using the Holm-Sidak method. *p < 0.05.

DOI: http://dx.doi.org/10.7554/eLife.06508.008

Figure 2—figure supplement 1. Nitrite and reactive oxygen production in PEMs following addition of inhibitors.

(A) PEM nitrite production following stimulation with LPS and IFN-γ and incubation with ROS (NAC) or NO (NAME) inhibitors as indicated. (B) Reactive oxygen production of PEMs after LPS or PMA stimulation with addition of indicated inhibitors.

DOI: http://dx.doi.org/10.7554/eLife.06508.009

Figure 2—figure supplement 2. Antimicrobial compounds (ROS and NO) enhance Perforin-2 mediated killing of M. smegmatis by PEM.

PEMs were isolated from either (A, C) wild-type, or (B, D) Perforin-2 knockout mice. □= MPEG1 (Perforin-2) +/+ PEMs, ■= MPEG1 (Perforin-2) −/− PEMs. Cells were incubated with NAC (blue line), NAME (green line), or mock (black line). The above graphs were conducted with eight biologic replicates and are representative of three experiments. Statistical analysis was performed utilizing multiple T-tests with correction for multiple comparisons using the Holm-Sidak method. *p < 0.05.

DOI: http://dx.doi.org/10.7554/eLife.06508.010