The Yersinia pestis Effector YopM Inhibits Pyrin Inflammasome Activation
Fig 2
LPS-primed BMDMs were infected with indicated Y. pestis strains for 6 hours; A) IL-1β, B) IL-18, and C) TNFα were measured in supernatants by ELISA 6 hrs p.i. (MOI 10); D) Total protein from LPS-primed BMDMs infected with indicated strains (combined cell lysate and supernatant) was separated by SDS-PAGE and analyzed by Western Blot for IL-1β and caspase-1. E) Cell death was assayed by LDH release in LPS-primed BMDMs infected with indicated strains at 6 hrs p.i. (MOI 10). F) Expression of Pyrin and Pro-IL-1β mRNA was measured by RT-PCR at 1, 3, 5, or 7 hours after addition of 100ng/mL LPS to WT BMDMs. G) WT C57Bl/6 (n = 12), Pyrin KO (n = 10), NLRP3 KO (n = 7) or IL-18 KO (n = 8) or H) WT (n = 9), Pyrin KO (n = 10) or caspase-1/11 KO (n = 8) mice were infected s.c. with Y. pestis KIM1001 ΔYopM/J (150 CFU) and monitored for survival up to 21 days. G, H): P value reflects comparison of WT vs Pyrin KO, NLRP3 vs Pyrin KO, WT vs IL-18 KO or WT vs caspase-1/11 KO. Figures are representative of three or more experiments, G, F are representative of two experiments. Shown is mean plus s.d. for triplicate wells. * p<0.05, **p<0.01, ***p<0.001.