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R&D Systems primary antibodies specific human il-1β

WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnFlaA 310–475 alone (LFnFlaA), LFnPrgJ alone, LFnYscF alone, PA+LFnFlaA 310–475 (FlaTox), PA+LFnPrgJ (PrgJTox), or PA+LFnYscF (YscFTox) for 6 hours (A, C). As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours (B, D). Release <t>of</t> <t>IL-1β</t> into the supernatant was measured by ELISA. ns–not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001 by Šídák’s multiple comparisons test (A), or by unpaired t-test (B), or by Dunnett’s multiple comparisons test (C, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Primary Antibodies Specific Human Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication"

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1009718

Figure Legend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnFlaA 310–475 alone (LFnFlaA), LFnPrgJ alone, LFnYscF alone, PA+LFnFlaA 310–475 (FlaTox), PA+LFnPrgJ (PrgJTox), or PA+LFnYscF (YscFTox) for 6 hours (A, C). As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours (B, D). Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001 by Šídák’s multiple comparisons test (A), or by unpaired t-test (B), or by Dunnett’s multiple comparisons test (C, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques Used: Clone Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Figure Legend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β (A, B) and IL-18 (C, D) into the supernatant were measured by ELISA. Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, *** p < 0.001, **** p < 0.0001 by Šídák’s multiple comparisons test (A, C) or Dunnett’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques Used: Clone Assay, Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

(A, C, E) WT, NAIP -/- , or NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. (B, D) As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours. (A–D) Release of IL-1β into the supernatant was measured by ELISA. (E) Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test (A, C) or by Šídák’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Figure Legend Snippet: (A, C, E) WT, NAIP -/- , or NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. (B, D) As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours. (A–D) Release of IL-1β into the supernatant was measured by ELISA. (E) Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test (A, C) or by Šídák’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques Used: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

WT, NAIP -/- (A–C) and NLRC4 -/- (D–F) THP-1 monocyte-derived macrophages were treated with siRNA targeting a control scrambled siRNA, siRNA targeting CASP4 (A, D) or CASP5 (B, E), or siRNA targeting both CASP4 and CASP5 (C, F) for 48 hours. Cells were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. As a control, cells were transfected with LPS. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test. Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Figure Legend Snippet: WT, NAIP -/- (A–C) and NLRC4 -/- (D–F) THP-1 monocyte-derived macrophages were treated with siRNA targeting a control scrambled siRNA, siRNA targeting CASP4 (A, D) or CASP5 (B, E), or siRNA targeting both CASP4 and CASP5 (C, F) for 48 hours. Cells were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. As a control, cells were transfected with LPS. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test. Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques Used: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Standard Deviation

WT, NAIP -/- (A, B), and NLRC4 -/- (C, D) THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20. Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. (A, C) CFU/well of bacteria at 6 hpi (B, D) Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. ns–not significant, *** p < 0.001, **** p < 0.0001 by Dunnett’s multiple comparisons test (A, C) or Tukey’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Figure Legend Snippet: WT, NAIP -/- (A, B), and NLRC4 -/- (C, D) THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20. Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. (A, C) CFU/well of bacteria at 6 hpi (B, D) Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. ns–not significant, *** p < 0.001, **** p < 0.0001 by Dunnett’s multiple comparisons test (A, C) or Tukey’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques Used: Derivative Assay, Infection, Bacteria, Standard Deviation

WT and NAIP -/- THP-1 monocyte-derived macrophages were seeded on glass coverslips and primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium expressing GFP at an MOI = 20. Cells were fixed at the indicated time points and stained for DAPI to label DNA (blue). The proportion of infected cells containing GFP-expressing Stm (green) and the number of bacteria per cell were scored by fluorescence microscopy. (A) Representative images from 6hpi are shown. Scale bar represents 10 μm. (B, C) Each small dot represents one infected cell. 150 infected cells were scored for each condition (50 infected cells per coverslip). Bars represent the mean from each condition. (B) Number of bacteria/cell at 2 hpi. (C) Number of bacteria/cell at 6 hpi. ns–not significant, **** p < 0.0001 by Tukey’s multiple comparisons test (B). Data shown are representative of at least three independent experiments.

Figure Legend Snippet: WT and NAIP -/- THP-1 monocyte-derived macrophages were seeded on glass coverslips and primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium expressing GFP at an MOI = 20. Cells were fixed at the indicated time points and stained for DAPI to label DNA (blue). The proportion of infected cells containing GFP-expressing Stm (green) and the number of bacteria per cell were scored by fluorescence microscopy. (A) Representative images from 6hpi are shown. Scale bar represents 10 μm. (B, C) Each small dot represents one infected cell. 150 infected cells were scored for each condition (50 infected cells per coverslip). Bars represent the mean from each condition. (B) Number of bacteria/cell at 2 hpi. (C) Number of bacteria/cell at 6 hpi. ns–not significant, **** p < 0.0001 by Tukey’s multiple comparisons test (B). Data shown are representative of at least three independent experiments.

Techniques Used: Derivative Assay, Infection, Expressing, Staining, Bacteria, Fluorescence, Microscopy

(A) Primary hMDMs from four healthy human donors were infected with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgJ (Lm + PrgJ), SsaI (Lm + SsaI), or SsaG (Lm + SsaG) for 16 hours at MOI = 5. Release of IL-18 into the supernatant was measured by ELISA. Each dot represents the mean of individual donors derived from triplicate wells. The grey bar represents the mean of all donors. (B) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were treated with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgI (Lm + PrgI), or Listeria expressing SsaG (Lm + SsaG) for 6 hours at MOI = 20. Release of IL-1β into the supernatant was measured by ELISA. (C) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnSsaG, or PA+LFnSsaG (SsaGTox) for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, *** p < 0.001, **** p < 0.0001 paired t-test (A) or by Dunnett’s multiple comparisons test (B, C). Data shown are representative of at least three independent experiments.

Figure Legend Snippet: (A) Primary hMDMs from four healthy human donors were infected with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgJ (Lm + PrgJ), SsaI (Lm + SsaI), or SsaG (Lm + SsaG) for 16 hours at MOI = 5. Release of IL-18 into the supernatant was measured by ELISA. Each dot represents the mean of individual donors derived from triplicate wells. The grey bar represents the mean of all donors. (B) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were treated with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgI (Lm + PrgI), or Listeria expressing SsaG (Lm + SsaG) for 6 hours at MOI = 20. Release of IL-1β into the supernatant was measured by ELISA. (C) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnSsaG, or PA+LFnSsaG (SsaGTox) for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, *** p < 0.001, **** p < 0.0001 paired t-test (A) or by Dunnett’s multiple comparisons test (B, C). Data shown are representative of at least three independent experiments.

Techniques Used: Infection, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay

WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or Δ prgIfliCfljB S . Typhimurium at an MOI = 20. (A, B) Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. (C, D) Release of IL-1β was measured at 24hpi by ELISA. ns–not significant, **** p < 0.0001 by Šídák’s multiple comparisons test.

Figure Legend Snippet: WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or Δ prgIfliCfljB S . Typhimurium at an MOI = 20. (A, B) Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. (C, D) Release of IL-1β was measured at 24hpi by ELISA. ns–not significant, **** p < 0.0001 by Šídák’s multiple comparisons test.

Techniques Used: Derivative Assay, Infection, Bacteria, Enzyme-linked Immunosorbent Assay

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Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnFlaA 310–475 alone (LFnFlaA), LFnPrgJ alone, LFnYscF alone, PA+LFnFlaA 310–475 (FlaTox), PA+LFnPrgJ (PrgJTox), or PA+LFnYscF (YscFTox) for 6 hours (A, C). As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours (B, D). Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001 by Šídák’s multiple comparisons test (A), or by unpaired t-test (B), or by Dunnett’s multiple comparisons test (C, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Clone Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β (A, B) and IL-18 (C, D) into the supernatant were measured by ELISA. Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, *** p < 0.001, **** p < 0.0001 by Šídák’s multiple comparisons test (A, C) or Dunnett’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques: Clone Assay, Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: (A, C, E) WT, NAIP -/- , or NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. (B, D) As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours. (A–D) Release of IL-1β into the supernatant was measured by ELISA. (E) Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test (A, C) or by Šídák’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- (A–C) and NLRC4 -/- (D–F) THP-1 monocyte-derived macrophages were treated with siRNA targeting a control scrambled siRNA, siRNA targeting CASP4 (A, D) or CASP5 (B, E), or siRNA targeting both CASP4 and CASP5 (C, F) for 48 hours. Cells were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. As a control, cells were transfected with LPS. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test. Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- (A, B), and NLRC4 -/- (C, D) THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20. Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. (A, C) CFU/well of bacteria at 6 hpi (B, D) Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. ns–not significant, *** p < 0.001, **** p < 0.0001 by Dunnett’s multiple comparisons test (A, C) or Tukey’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Techniques: Derivative Assay, Infection, Bacteria, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT and NAIP -/- THP-1 monocyte-derived macrophages were seeded on glass coverslips and primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium expressing GFP at an MOI = 20. Cells were fixed at the indicated time points and stained for DAPI to label DNA (blue). The proportion of infected cells containing GFP-expressing Stm (green) and the number of bacteria per cell were scored by fluorescence microscopy. (A) Representative images from 6hpi are shown. Scale bar represents 10 μm. (B, C) Each small dot represents one infected cell. 150 infected cells were scored for each condition (50 infected cells per coverslip). Bars represent the mean from each condition. (B) Number of bacteria/cell at 2 hpi. (C) Number of bacteria/cell at 6 hpi. ns–not significant, **** p < 0.0001 by Tukey’s multiple comparisons test (B). Data shown are representative of at least three independent experiments.

Techniques: Derivative Assay, Infection, Expressing, Staining, Bacteria, Fluorescence, Microscopy

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: (A) Primary hMDMs from four healthy human donors were infected with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgJ (Lm + PrgJ), SsaI (Lm + SsaI), or SsaG (Lm + SsaG) for 16 hours at MOI = 5. Release of IL-18 into the supernatant was measured by ELISA. Each dot represents the mean of individual donors derived from triplicate wells. The grey bar represents the mean of all donors. (B) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were treated with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgI (Lm + PrgI), or Listeria expressing SsaG (Lm + SsaG) for 6 hours at MOI = 20. Release of IL-1β into the supernatant was measured by ELISA. (C) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnSsaG, or PA+LFnSsaG (SsaGTox) for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, *** p < 0.001, **** p < 0.0001 paired t-test (A) or by Dunnett’s multiple comparisons test (B, C). Data shown are representative of at least three independent experiments.

Techniques: Infection, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or Δ prgIfliCfljB S . Typhimurium at an MOI = 20. (A, B) Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. (C, D) Release of IL-1β was measured at 24hpi by ELISA. ns–not significant, **** p < 0.0001 by Šídák’s multiple comparisons test.

Techniques: Derivative Assay, Infection, Bacteria, Enzyme-linked Immunosorbent Assay

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