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: 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: Genetics and Molecular Research

Article Title: Protective effects of bifidobacteria on intestines in newborn rats with necrotizing enterocolitis and its regulation on TLR2 and TLR4

doi: 10.4238/2015.september.28.2

Figure Lengend Snippet: Figure 2. Immunohistochemistry can detect the expression level of TLR2 in neonatal intestinal tissues of rats in each group (200X). TLR2 was expressed in each group, and was more obvious in crypts. There was weakly positive staining on the top of intestinal epithelial cytoplasm in groups C (c), D (d), and E (e), while there was strong positive expression on villi and crypts in groups B (b) and A (a).

Article Snippet: The TLR2 immunohistochemistry primary antibody (rabbit anti-rat) was purchased from GenWay Biotech, Inc. (San Diego, CA, USA).

Techniques: Immunohistochemistry, Expressing, Staining

Journal: Genetics and Molecular Research

Article Title: Protective effects of bifidobacteria on intestines in newborn rats with necrotizing enterocolitis and its regulation on TLR2 and TLR4

doi: 10.4238/2015.september.28.2

Figure Lengend Snippet: Figure 3. Immunohistochemistry can detect the expression level of TLR4 in neonatal intestinal tissues of rats in each group (200X). TLR4 in groups C (c), D (d) and E (e) was only weak expressed in rat intestinal epithelial cells, and not expressed in the submucosa. There was an enhanced and increased expression in group A (a), while in group B (b) the expression was significantly increased, showing diffuse or granular distribution.

Article Snippet: The TLR2 immunohistochemistry primary antibody (rabbit anti-rat) was purchased from GenWay Biotech, Inc. (San Diego, CA, USA).

Techniques: Immunohistochemistry, Expressing

Journal: Genetics and Molecular Research

Article Title: Protective effects of bifidobacteria on intestines in newborn rats with necrotizing enterocolitis and its regulation on TLR2 and TLR4

doi: 10.4238/2015.september.28.2

Figure Lengend Snippet: Figure 4. Immunohistochemistry can detect the expression level of NF-кB p65 in neonatal intestinal tissues of rats in each group (200X). There was no nuclear staining of NF-кB p65 in intestinal villi of Group E (e) rats. In groups C (c) and D (d), some nuclei stained can be seen. Nuclear staining increased significantly in group A (a), while Group B (b) showed a large nuclear staining.

Article Snippet: The TLR2 immunohistochemistry primary antibody (rabbit anti-rat) was purchased from GenWay Biotech, Inc. (San Diego, CA, USA).

Techniques: Immunohistochemistry, Expressing, Staining

Journal: Parasites & vectors

Article Title: Pentatrichomonas hominis induces extracellular traps formation of macrophages via the TLR2/NADPH/PAD4 pathway.

doi: 10.1186/s13071-025-06840-w

Figure Lengend Snippet: Fig. 6 TLR2 was involved in the formation of METs induced by P. hominis trophozoites. a, To detect the expression of Tlr2, macrophages (7.5 × 105 cells/ml) were stimulated with P. hominis trophozoites or zymosan for 30, 60, 90, and 120 min. The internal reference gene used was β-actin. b, Macrophages were pretreated with the TLR2 inhibitor C29 prior to P. hominis trophozoite stimulation. The dsDNA produced by METs in supernatants was stained with PicoGreen and detected. The macrophages stimulated with zymosan were used as positive controls, and the unstimulated macrophages were used as negative controls. Bars represent the mean ± SD for three experiments. ns, not significant. *P < 0.05, ***P < 0.001, ****P < 0.0001

Article Snippet: In parallel experiments, macrophages were pretreated with 20 μM of NADPH oxidase inhibitor (diphenylene iodonium [DPI], Merck, D2926, USA), 100 μM of MPO inhibitor (4-aminobenzoic acid hydrazide [4-ABAH], Selleck, S9874, China), 50 μM of extracellular regulated protein kinase 1/2 (ERK1/2) inhibitor (U0126, Selleck, S1102, China), 10 μM of p38 MAPK inhibitor (SB202190, Selleck, S1077, China), 100 μM of TLR2 inhibitor (TLR2-IN-C29, Selleck, S6597, China), 100 μM of SOCE inhibitor (2-aminoethyl diphenylborinate [2-APB], Selleck, S6657, China), 10 μM of PAD4 inhibitor (GSK484 hydrochloride, MedChemExpress, HY-100514, USA), and 1 μM of cytochalasin D (Cyt D, APExBio, 22144-77- 0, USA) at 37 °C and 5% CO2 for 30 min. DNase I in 90 U/well (Thermo Scientific, EN0523, USA) was used to digest the MET structure for 15 min before the end of coincubation.

Techniques: Expressing, Produced, Staining

Journal: Parasites & vectors

Article Title: Pentatrichomonas hominis induces extracellular traps formation of macrophages via the TLR2/NADPH/PAD4 pathway.

doi: 10.1186/s13071-025-06840-w

Figure Lengend Snippet: Fig. 7 ERK1/2, p38 MAPK signaling pathway, PAD4 and SOCE were involved in P. hominis-induced MET formation, TLR2 and p38 MAPK are involved in the induction of ROS. The macrophages were pretreated with: a, an ERK1/2 inhibitor (U0126), b, a p38 MAPK inhibitor (SB202190), c, a PAD4 inhibitor (GSK484), and d, a SOCE inhibitor (2-APB) for 30 min and then coincubated with P. hominis. The dsDNA produced by METs in supernatants was stained with PicoGreen and detected. e, Macrophages were pretreated with C29 and SB202190; ROS production was detected via DCFH-DA. Zymosan-stimulated macrophages were used as a positive control, and unstimulated macrophages were used as a negative control. Bars represent the mean ± SD for three experiments. ns, not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: In parallel experiments, macrophages were pretreated with 20 μM of NADPH oxidase inhibitor (diphenylene iodonium [DPI], Merck, D2926, USA), 100 μM of MPO inhibitor (4-aminobenzoic acid hydrazide [4-ABAH], Selleck, S9874, China), 50 μM of extracellular regulated protein kinase 1/2 (ERK1/2) inhibitor (U0126, Selleck, S1102, China), 10 μM of p38 MAPK inhibitor (SB202190, Selleck, S1077, China), 100 μM of TLR2 inhibitor (TLR2-IN-C29, Selleck, S6597, China), 100 μM of SOCE inhibitor (2-aminoethyl diphenylborinate [2-APB], Selleck, S6657, China), 10 μM of PAD4 inhibitor (GSK484 hydrochloride, MedChemExpress, HY-100514, USA), and 1 μM of cytochalasin D (Cyt D, APExBio, 22144-77- 0, USA) at 37 °C and 5% CO2 for 30 min. DNase I in 90 U/well (Thermo Scientific, EN0523, USA) was used to digest the MET structure for 15 min before the end of coincubation.

Techniques: Produced, Staining, Positive Control, Negative Control

Journal: Cells

Article Title: Role of TLR4 Receptor Complex in the Regulation of the Innate Immune Response by Fibronectin

doi: 10.3390/cells9010216

Figure Lengend Snippet: TLR4 mediates IL-8 expression in response to FnIII-1c and LPS in dermal fibroblasts. Monolayers of human dermal fibroblasts in 10% FBS/DMEM were treated for 24 h with ( A ) FnIII-1c or FnIII-13 (1-20 µg/mL), ( B ) LPS (1-100 ng/mL), ( C ) LPS (100 ng/mL) or FnIII-1c (10 µM) in the presence of the designated amounts of blocking antibody to TLR4 or TLR2. IgG served as control. ( D ) TNF-α (25 ng/mL), LPS (100 ng/mL) or FnIII-1c (10 µM) in the presence of increasing amounts of the TLR4 inhibitor, TAK-242. The wells without antibodies ( C ) or inhibitors ( D ) were set as 100%. IL-8 concentration in conditioned medium was determined by ELISA. The data represent the mean ± S.E. of triplicate assays from three separate experiments.

Article Snippet: Recombinant human CD14, human TNF-α, human IL-1α, anti-human MD-2 antibody, and neutralizing antibodies: anti-human CD14, anti-human TLR2 and anti-human TLR4 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Blocking Assay, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay