lnye01000004 1  (ATCC)

Journal: PLoS Pathogens

Article Title: Inhibition of caspase-1 or gasdermin-D enable caspase-8 activation in the Naip5/NLRC4/ASC inflammasome

doi: 10.1371/journal.ppat.1006502

Figure Lengend Snippet: (A-D) Bone marrow-derived macrophages (BMDMs) from C57BL/6 (open circles), Nlrc4 -/- (closed squares), Casp1/11 -/- (closed triangles) and Asc/Casp1/11 -/- (open triangles) mice were infected with motility-deficient mutants expressing flagellin ( fliI - , A), with flagellin-deficient mutants ( flaA - , B), L . gratiana (C) or with L . micdadei (D) at a MOI of 10. The cells were incubated for 24, 48 and 72 hours for CFU determination. Data show the average ± SD of triplicate wells. * , P <0.05 compared with Casp-1/11 -/- BMDMs. # , P <0.05 compared with C57BL/6 BMDMs, ANOVA. (E-I) BMDMs from Casp1/11 -/- and Asc/Casp1/11 -/- mice were transduced with a retrovirus encoding shRNA sequences to target caspase-8 (Seq1, Seq2) and a non-target shRNA sequence (NT). (E) Caspase-8 silencing was confirmed by western blot analysis. Cell lysates were separated by SDS-PAGE, blotted and probed with anti-caspase-8 (pro-caspase-8 p55) and anti-α-actin. (F-I) Transduced Casp1/11 -/- (F, H) and Asc/Casp1/11 -/- (G, I) BMDMs were infected with fliI - (F, G) or flaA - (H, I) at a MOI of 10 and incubated for 24, 48 and 72 hours for CFU determination. Data show the average ± SD of triplicate wells. * , P <0.05 compared with NT shRNA, ANOVA. NT, non-target shRNA. Data are presented for one representative experiment of four (A), two (B-D) and one (F-I) experiments performed with similar results.

Article Snippet: L . micdadei (ATCC 33218) and L . gratiana (ATCC 49413) were used to generate streptomycin-resistant strains.

Techniques: Derivative Assay, Infection, Expressing, Incubation, Transduction, shRNA, Sequencing, Western Blot, SDS Page

Journal: PLoS Pathogens

Article Title: Inhibition of caspase-1 or gasdermin-D enable caspase-8 activation in the Naip5/NLRC4/ASC inflammasome

doi: 10.1371/journal.ppat.1006502

Figure Lengend Snippet: C57BL/6 (open circles), Nlrc4 -/- (closed squares), Casp1/11 -/- (closed triangles) and Asc/Casp1/11 -/- (open triangles) mice were infected with motility-deficient L . pneumophila mutants expressing flagellin ( fliI - , A, D), with flagellin-deficient L . pneumophila ( flaA - , D), with L . gratiana (B) or with L . micdadei (C) at a dose of 10 5 bacteria/mice. The mice were euthanized at 4, 48 or 72 hours after infection. Dilutions of the lung homogenates were added to charcoal-yeast extract agar plates for colony-forming unit determination. Each dot represents a single animal, and the horizontal lines represent the averages. * , P <0.05. # , indicates P <0.05 compared with C57BL/6, Mann Whitney test. Data are presented for one representative experiment of five (A), one (B-C) and two (D) experiments performed with similar results.

Article Snippet: L . micdadei (ATCC 33218) and L . gratiana (ATCC 49413) were used to generate streptomycin-resistant strains.

Techniques: Infection, Expressing, Bacteria, MANN-WHITNEY