Journal: BMB Reports

Article Title: Diesel exhaust particles disrupt blood–retina barrier integrity via TLR2 and TLR4 activation

doi: 10.5483/BMBRep.2025-0013

Figure Lengend Snippet: Expression levels of TLR2/TLR4 and TNF-α/IL-1β mRNA in U937 macrophages exposed to DEPs with TLR2 (C29) or TLR4 (TAK242) inhibitors. (A) Schematic representation of the experimental procedure. (B, C) TLR2 and TLR4 expression levels. Compared with those in the control, TLR2 (B) and TLR4 (C) levels in U937 macrophages were significantly elevated following DEP exposure. TLR2 and TLR4 expression levels were significantly restored following treatment with C29 and TAK242 before DEP exposure, respectively. (D, E) TNF-α and IL-1β mRNA expression. TNF-α (D) and IL-1β (E) mRNA levels were significantly lower in U937 macrophages treated with C29 or TAK242 before DEP exposure than in cells that received DEP exposure alone without TLR inhibitors. *P < 0.05 vs. control, † P < 0.05 vs. DEP.

Article Snippet: The membranes were blocked with 5% skim milk in PBS containing 0.1% Tween 20 (PBSTw) for 1 h and were then incubated with the following primary antibodies: anti-mouse TLR2 (1:100, Santa Cruz Biotechnology, TX, USA), anti-mouse TLR4 (1:100, Santa Cruz Biotechnology), anti-mouse claudin-5 (1:500, Invitrogen), anti-rabbit ZO-1 (1:1000, Invitrogen) and anti-mouse actin (JLA20, 1:10, DSHB, IA, USA).

Techniques: Expressing, Control

Journal: PloS one

Article Title: Immunogenecity of modified alkane polymers is mediated through TLR1/2 activation.

doi: 10.1371/journal.pone.0002438

Figure Lengend Snippet: Figure 5. Modified alkane polymers induce activation of TLR-1 and TLR-2 signaling pathways. a) Luciferase activity expressed by human TLR1/2, TLR2, TLR3 and TLR4 stable HEK 3T3 transfectant (pNF-kB-LUC Stratagene). Cells were left untreated or treated for a different time period with 50 mg/ml of unPE, mPE or pre and post implant PE and respective positive controls; PGN(10 mg/ml) for TLR1/2 and TLR2, Poly (I:C)(1 mg/ml) for TLR3 and LPS (10 mg/ml) for TLR4. doi:10.1371/journal.pone.0002438.g005

Article Snippet: Similar experiments were conducted in presence of mouse anti human TLR2 mAb (clone 383936 R&D Systems) which is known to prevent ligand access to the TLR2 binding groove.

Techniques: Modification, Activation Assay, Protein-Protein interactions, Luciferase, Activity Assay, Transfection

Journal: PloS one

Article Title: Immunogenecity of modified alkane polymers is mediated through TLR1/2 activation.

doi: 10.1371/journal.pone.0002438

Figure Lengend Snippet: Figure 6. Direct binding of oxidized alkane polymers to soluble TLR-2 molecules. a) Left panels; fluorescence emission scans collected for free soluble TLR2 and TLR2 in complex with two different concentrations (exponential and plateau) of each analyzed polymer. Central panels; normalized fluorescence data (DF) for each concentration point as a function of the free ligand concentration. Right panels; fluorescence emission scans collected for free soluble TLR2 and TLR2 in complex with each analyzed polymer at two different concentrations (exponential and plateau) in presence of an anti TLR2 mAb known to block the TLR2 binding groove (stoichiometric ratio 2:1 Ab to soluble TLR2 receptor). b) Comparison of the fluorescence emission scans collected for soluble TLR2, mPE and unPE. doi:10.1371/journal.pone.0002438.g006

Article Snippet: Similar experiments were conducted in presence of mouse anti human TLR2 mAb (clone 383936 R&D Systems) which is known to prevent ligand access to the TLR2 binding groove.

Techniques: Binding Assay, Fluorescence, Polymer, Concentration Assay, Blocking Assay, Comparison

Journal: PLOS ONE

Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis

doi: 10.1371/journal.pone.0302813

Figure Lengend Snippet: Baseline characteristics and comparison stratified by Toll-like receptor (TLR) 2 cytoplasmic intensity in primary tumors and lymph node metastases.

Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337), TLR2 for 60 minutes (diluted 1:500, Rockland 600-401-956), TLR3 for 120 minutes (diluted 1:30, Novus NBP2-24875), TLR4 for 60 minutes (diluted 1:1000, Abnova H00007099-M02), TLR5 for overnight in +4°C (diluted 1:75, Novus NBP2-24787), TLR6 for 60 minutes (diluted 1:750, Abnova PAB 3555), TLR7 for 60 minutes (diluted 1:500, Novus NB100-56682), TLR8 for 60 minutes (diluted 1:850, Novus NBP2-24917) and TLR9 for 60 minutes (diluted 1:300, Novus NBP2-24729).

Techniques: Comparison

Journal: PLOS ONE

Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis

doi: 10.1371/journal.pone.0302813

Figure Lengend Snippet: Immunohistochemical staining examples of Toll-like receptors (TLRs) 1, 2, 4, 5, 6, 7, 8 and 9 in representative small bowel neuroendocrine tumor samples showing ( A ) High TLR1 cytoplasmic intensity, ( B ) intermediate TLR2 cytoplasmic intensity, ( C ) intermediate TLR4 cytoplasmic intensity, ( D ) high TLR5 cytoplasmic and nucleic intensity, ( E ) Intermediate TLR6 cytoplasmic intensity, ( F ) intermediate TLR7 cytoplasmic intensity, ( G ) high TLR8 cytoplasmic intensity and ( H ) intermediate TLR9 cytoplasmic intensity in x20 magnification. The scale bar length is 50 μm (bottom left corner). Arrows indicate TLR-positive tumor cell islets.

Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337), TLR2 for 60 minutes (diluted 1:500, Rockland 600-401-956), TLR3 for 120 minutes (diluted 1:30, Novus NBP2-24875), TLR4 for 60 minutes (diluted 1:1000, Abnova H00007099-M02), TLR5 for overnight in +4°C (diluted 1:75, Novus NBP2-24787), TLR6 for 60 minutes (diluted 1:750, Abnova PAB 3555), TLR7 for 60 minutes (diluted 1:500, Novus NB100-56682), TLR8 for 60 minutes (diluted 1:850, Novus NBP2-24917) and TLR9 for 60 minutes (diluted 1:300, Novus NBP2-24729).

Techniques: Immunohistochemical staining, Staining

Journal: PLOS ONE

Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis

doi: 10.1371/journal.pone.0302813

Figure Lengend Snippet: Median Toll-like receptor (TLR) staining intensity in primary small bowel neuroendocrine tumors and lymph node metastases.

Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337), TLR2 for 60 minutes (diluted 1:500, Rockland 600-401-956), TLR3 for 120 minutes (diluted 1:30, Novus NBP2-24875), TLR4 for 60 minutes (diluted 1:1000, Abnova H00007099-M02), TLR5 for overnight in +4°C (diluted 1:75, Novus NBP2-24787), TLR6 for 60 minutes (diluted 1:750, Abnova PAB 3555), TLR7 for 60 minutes (diluted 1:500, Novus NB100-56682), TLR8 for 60 minutes (diluted 1:850, Novus NBP2-24917) and TLR9 for 60 minutes (diluted 1:300, Novus NBP2-24729).

Techniques: Staining

Journal: PLOS ONE

Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis

doi: 10.1371/journal.pone.0302813

Figure Lengend Snippet: Disease-specific survival rates based on Toll-like receptor (TLR) 1, 2, 4, 5, 6, 7, 8, and 9 cytoplasmic staining intensity and TLR5 nucleic staining intensity in both primary tumors and lymph node metastases.

Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337), TLR2 for 60 minutes (diluted 1:500, Rockland 600-401-956), TLR3 for 120 minutes (diluted 1:30, Novus NBP2-24875), TLR4 for 60 minutes (diluted 1:1000, Abnova H00007099-M02), TLR5 for overnight in +4°C (diluted 1:75, Novus NBP2-24787), TLR6 for 60 minutes (diluted 1:750, Abnova PAB 3555), TLR7 for 60 minutes (diluted 1:500, Novus NB100-56682), TLR8 for 60 minutes (diluted 1:850, Novus NBP2-24917) and TLR9 for 60 minutes (diluted 1:300, Novus NBP2-24729).

Techniques: Staining

Journal: Frontiers in microbiology

Article Title: Human herpesviruses-encoded dUTPases: a family of proteins that modulate dendritic cell function and innate immunity.

doi: 10.3389/fmicb.2014.00504

Figure Lengend Snippet: FIGURE 3 | Activation of NF-κB by herpesviruses-encoded dUTPases is weaker in TLR2/TLR6-HEK293 than in TLR2/TLR1-HEK293 expressing cells. Herpesviruses-encoded dUTPases mediated activation of NF-κB in (A) TLR2-HEK293 and (B) TLR2/TLR6-HEK293 cell lines. Cells were transiently transfected with NF-κB luciferase and pRL-TK reporter plasmids and co-transfected with empty vector, as described in Materials and Methods. Empty vector was used as a carrier to keep the total amount of transfected-DNA constant. After 24–36 h, cells were treated with various herpesviruses-encoded dUTPases (10 μg/ml) or left untreated for 8 h, and NF-κB luciferase levels were measured. Zymosan (10 μg/ml) and FSL-1 (0.1 μg/ml) were used as positive controls for TLR2 and TLR6 activation, respectively. Data were normalized for transfection efficiency by measuring Renilla luciferase activity and expressed as the mean fold induction ± SD relative to control levels. Values represent the average of three independent experiments.

Article Snippet: IgG2a Isotype control monoclonal antibody was purchased from eBioscience (San Diego, California) and anti-TLR2 (clone TL2.1) monoclonal antibody was purchased from Imgenex (San Diego, California).

Techniques: Activation Assay, Expressing, Transfection, Luciferase, Plasmid Preparation, Activity Assay, Control

Journal: Frontiers in microbiology

Article Title: Human herpesviruses-encoded dUTPases: a family of proteins that modulate dendritic cell function and innate immunity.

doi: 10.3389/fmicb.2014.00504

Figure Lengend Snippet: FIGURE 4 | Herpesviruses-encoded dUTPases signal through TLR2/TLR1 and MyD88. TLR2- HEK293 cells were transiently transfected with NF-κB luciferase and pRL-TK reporter plasmids and co-transfected with pCMV-TLR1 and either TLR2DN, a vector that expresses a dominant-negative form of TLR2, or empty vector (0.3 μg) (A). After 24–36 h, cells were treated with the dUTPases (10 μg/ml) encoded by HHV-6A, HHV-8 and VZV or left untreated for 8 h, and luciferase reporter gene activity was measured. Pam3Csk4 (0.1 μg/ml) was used as a positive control for TLR2/TLR1 activation and nuclear human dUTPase was used as a control protein. (B) Anti-TLR2 blocking Ab inhibits NF-κB activation by the herpesviruses-encoded dUTPases. TLR2-HEK293 cells were transiently (Continued)

Article Snippet: IgG2a Isotype control monoclonal antibody was purchased from eBioscience (San Diego, California) and anti-TLR2 (clone TL2.1) monoclonal antibody was purchased from Imgenex (San Diego, California).

Techniques: Transfection, Luciferase, Plasmid Preparation, Dominant Negative Mutation, Activity Assay, Positive Control, Activation Assay, Control, Blocking Assay

Journal: International Journal of Molecular Sciences

Article Title: PAUF Induces Migration of Human Pancreatic Cancer Cells Exclusively via the TLR4/MyD88/NF-κB Signaling Pathway

doi: 10.3390/ijms231911414

Figure Lengend Snippet: The primer sequences of the genes for mRNA expression quantification by qPCR.

Article Snippet: Antibodies against TLR2 (cat# AF2616), TLR4 (cat# AF1478), and Goat IgG-phycoerythrin (IgG-PE, cat# F0107) were purchased from R&D Systems, Inc (Minneapolis, MN, USA). eFluor 450 conjugated mouse anti-PDL1 antibody (cat# 48-5983-42) and eFluor 450 conjugated mouse IgG1, kappa isotype control (cat# 48-4714-82) from Invitrogen (Waltham, MA, USA) was used for detection of surface PD-L1 expression in PC cells.

Techniques: Expressing