piκb α  (Cell Signaling Technology Inc)

Journal: Frontiers in Physiology

Article Title: Expression of Chitotriosidase in Macrophages Modulates Atherosclerotic Plaque Formation in Hyperlipidemic Mice

doi: 10.3389/fphys.2020.00714

Figure Lengend Snippet: List of antibodies used in Western Blot and IHC.

Article Snippet: PIκB-α (Ser32) , 2859 , Cell signaling , 1:1,000.

Techniques: Western Blot

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: Effect of co-culture of A. actinomycetemcomitans strain Y4 (Aa- MOI 1:100) with U937 macrophages for 1, 2, 3, 4 and 8 h on the relative transcription of TLR4, TLR2, NLRP3, NOD1, NOD2 IL-1β, IL-18 and TNF-α detected by real-time PCR and expressed in fold-change in comparison with control. Control consisted of infected cells at 0 h. Transcription of target gene was normalized according to mRNA levels of GAPDH; data are shown as fold-change ± SD representative of three independent experiments.

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Real-time Polymerase Chain Reaction, Comparison, Control, Infection, Expressing

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: Effect of co-culture of A. actinomycetemcomitans strain Y4 (Multiplicity of infection (MOI) 1:100) with U937 macrophages ( A ) or HGECs ( B ) for 1, 2, 3, 4 and 8 h on the levels of IL-1β and TNF-α levels in cell supernatants. Control consisted of infected cells at 0 h. Data (pg cytokine/mL) are presented as mean ± SD representative of three independent experiments. * Statistically significant difference in comparison with control (ANOVA–Tukey’s, p < 0.05).

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Co-Culture Assay, Infection, Control, Comparison

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: Different pathways are activated by A. actinomycetemcomitans in macrophages and HGECs. Western blot was used to evaluate the phosphorylation of ERK1/2, 4EBP-1, cFos and AKT in U937 macrophages ( A ) and in HGECs ( B ) after infection with A. actinomycetemcomitans strain Y4 (MOI 1:100) at different time points. GAPDH was used as the control. The data shown are representative of three independent experiments.

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Western Blot, Phospho-proteomics, Infection, Control

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: Western blot image showing increased levels of phosphorylated IκB (plkB-α), (indicative of NF-κB activation), decreased levels of procaspase-1 and increased levels of cleaved caspase-1 (indicative of inflammasome activation in U937 macrophages) after co-culture of A. actinomycetemcomitans strain Y4 (MOI 1:100) at different time points. GAPDH was used as the control. The data shown are representative of three independent experiments.

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Western Blot, Activation Assay, Co-Culture Assay, Control

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: Effect of siNLRP3 silencing in A. actinomycetemcomitans Y4-infected U937 macrophages (MOI 1:100, 24 h of co-culture) on the relative expression of NLRP3 ( A ), IL-1β ( B ) , IL-18 ( C ) and TNF-α ( D ) detected by real time PCR. Scramble: Scramble control without A. actinomycetemcomitans (pool of nontargeting Sirna) ; negative control: Cells without A. actinomycetemcomitans ; control: Positive control: Cells with A. actinomycetemcomitans without NLRP3 silencing; siNLRP3: Cells with silencing of NLRP3 and co-culture with A. actinomycetemcomitans . * Statistically significant difference in comparison with electroporated cells with A. actinomycetemcomitans and scramble control (ANOVA–Tukey’s, p < 0.05). Data shown in fold-change relative to the scramble are presented as mean ± SD representative of three independent experiments.

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Infection, Co-Culture Assay, Expressing, Real-time Polymerase Chain Reaction, Control, Negative Control, Positive Control, Comparison

Journal: Pathogens

Article Title: Distinct Signaling Pathways Between Human Macrophages and Primary Gingival Epithelial Cells by Aggregatibacter actinomycetemcomitans

doi: 10.3390/pathogens9040248

Figure Lengend Snippet: U937 macrophage response after co-culture with A. actinomycetemcomitans Y4. Arrows mean up- or downregulation of gene transcription for TLR4, TLR2, NOD2, NLRP3, Pro - IL-1β, Pro-IL-18 and TNF-α, and for phosphorylation of 4EBP1, AKT, pERK1/2, IκB-α and caspase-1. A. actinomycetemcomitans was able to induce an inflammatory response and inflammasome activation in macrophages. Adapted from Qiagen’s website ( https://www.qiagen.com/br/shop/genes-and-pathways/pathway-central/?q =).

Article Snippet: In U937 cells, pIκB-α (Ser32) (Cell Signaling, Danvers, MA, USA), caspase-1 p10 (C20) and cleaved caspase p10 were also evaluated.

Techniques: Co-Culture Assay, Phospho-proteomics, Activation Assay