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Image Search Results
Journal: PLoS Pathogens
Article Title: MyD88 and IL-1R signaling drive antibacterial immunity and osteoclast-driven bone loss during Staphylococcus aureus osteomyelitis
doi: 10.1371/journal.ppat.1007744
Figure Lengend Snippet: (A) Osteoclast precursors were generated from primary WT, Myd88 -/- , and Il1r1 -/- BMMs by treatment with 35 ng/mL RANKL treatment for 48 hours. After washing 2X with PBS, fresh media was replenished, and cells were stimulated with vehicle (1% casamino acid-supplemented RPMI) or toxin-deficient S. aureus supernatant (Δ psm ) in the absence of ongoing RANKL treatment. All cells were TRAP stained in vitro at day 6 and TRAP + multinucleated cells were manually quantified using OsteoMeasure software ( n = 3 wells per condition, representative of duplicate experiments). (B) WT and Il1r1 -/- enriched BMMs were stimulated with 35 ng/mL exogenous RANKL for 2 days before stimulating with Δ psm supernatants for four days. In order to determine the contribution of IL-1 signaling in this assay, vehicle (0.1% low endotoxin BSA) or recombinant murine IL-1ra (1 μg/mL) were added during the 2 days of RANKL pre-commitment (BSA, black bars; IL-1ra, grey bars) or during the 4 days of Δ psm supernatant stimulation (indicated by + under each bar). The top line of each bar represents the mean and error bars represent standard deviation. A repeated measures two-way ANOVA with Dunnett’s multiple comparisons test was performed on cell counts from each genotype, comparing each dose of Δ psm supernatant to the vehicle control, where * p < 0.05, *** p < 0.001, **** p < 0.0001, and if not designated by asterisks is non-significant. A repeated measures two-way ANOVA with Tukey’s multiple comparisons test was performed to compare Myd88 -/- and Il1r1 -/- TRAP + multinucleated cell counts to WT counts within each Δ psm supernatant dose, where # p < 0.05, † p < 0.001, and ‡ p < 0.0001. A three-way ANOVA with Tukey’s multiple comparisons test compared the effects of genotype, IL-1ra treatment during RANKL pre-commitment, and IL-1ra treatment during Δ psm supernatant stimulation. Statistical comparisons to WT cells pre-committed with RANKL + BSA and Δ psm supernatant + BSA (first black bar from the left) are reported as @ p < 0.05, or compared to WT cells pre-committed with RANKL + BSA and Δ psm supernatant + IL-1ra (second black bar from the left) are reported as § p < 0.05. Comparisons between all RANKL + IL-1ra pre-commitment conditions (grey bars) were non-significant.
Article Snippet: To test the specific role of IL-1R inhibition on S . aureus -enhanced osteoclast differentiation, osteoclastogenesis assays in WT and Il1r1 -/- cells were conducted with the addition of a vehicle control (0.1% low endotoxin BSA) or 1 μg/mL
Techniques: Generated, Staining, In Vitro, Software, Recombinant, Standard Deviation, Control
Journal:
Article Title: Differential expression of immunoregulatory genes in monocytes in response to Porphyromonas gingivalis and Escherichia coli lipopolysaccharide
doi: 10.1111/j.1365-2249.2009.03920.x
Figure Lengend Snippet: Primers and probes for the quantification of gene expression using Taq man low-density arrays (TLDAs).
Article Snippet: RNA polymerase II (Hs00172187_ml) was used as an endogenous control. table ft1 table-wrap mode="anchored" t5 caption a7 Inflammatory mediator Primers and probe assay ID IL-1α (interleukin-1 alpha) Hs00174092_m1 IL-1β (interleukin-1 beta) Hs00174097_m1 IL-Ra (interleukin-1 receptor antagonist)
Techniques: Gene Expression, Activation Assay, Control