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: Neural Plasticity

Article Title: Toll-Like Receptor 2 Attenuates Traumatic Brain Injury-Induced Neural Stem Cell Proliferation in Dentate Gyrus of Rats

doi: 10.1155/2020/9814978

Figure Lengend Snippet: Immunofluorescence (IF) microphotographs in the dentate gyrus (DG) of the DG in sham and TBI groups at different time points. (a) BrdU-labelled NSCs (red fluor), expression of TLR2 (green fluor), cell nuclei (blue fluor), and BrdU + /TLR2 + /DAPI + cells indicated that TLR2 expression in labelled proliferating cells was possible NSCs; (b) BrdU + cells showed the proliferation of NSCs in the DG during different time points posttrauma. BrdU + cells were more in the TBI group than that in the sham group ( ∗ p < 0.05), and numbers of these cells were significantly different among different time points posttrauma ( # p < 0.05); (c) numbers of BrdU + /TLR2 + cells indicated that the expression of TLR2 was quite different in proliferating cells of the DG among different time points posttrauma. There were more BrdU + cells in the TBI group than that in the sham group ( ∗ p < 0.05), and the numbers of these cells are obviously different among different time points posttrauma ( # p < 0.05). Scale bar: 50 μ m; data is shown as mean ± SEM.

Article Snippet: Primary antibodies were used as follows: mouse anti-TLR2 antibody (1 : 500, Biorbyt, orb191498, San Francisco, CA, USA) and mouse anti- β -actin antibody (1 : 3000, TDY BIOTECH, TDY041, Beijing, China) overnight at 4°C.

Techniques: Immunofluorescence, Expressing

Journal: Neural Plasticity

Article Title: Toll-Like Receptor 2 Attenuates Traumatic Brain Injury-Induced Neural Stem Cell Proliferation in Dentate Gyrus of Rats

doi: 10.1155/2020/9814978

Figure Lengend Snippet: IF in the DG of the TBI group (mouse brain got from 3 days posttrauma). (a) BrdU (red), nestin (green), and DAPI (blue), respectively, exhibited proliferating cells, NSCs, and cell nuclei in the DG. Merged pictures of BrdU + /nestin + /DAPI + showed NSCs (the percentage of NSCs in proliferating cells was 84.30% ± 6.54%); scale bar: 50 μ m; data were expressed as mean ± SEM. (b) Nestin (green), TLR2 (red), and DAPI (blue), respectively, exhibited NSCs, TLR2 expression, and cell nuclei in the DG. Merged pictures of nestin + /TLR2 + /DAPI + showed the expression of TLR2 on NSCs. Scale bar: 50 μ m; data were expressed as mean ± SEM.

Article Snippet: Primary antibodies were used as follows: mouse anti-TLR2 antibody (1 : 500, Biorbyt, orb191498, San Francisco, CA, USA) and mouse anti- β -actin antibody (1 : 3000, TDY BIOTECH, TDY041, Beijing, China) overnight at 4°C.

Techniques: Expressing

Journal: Neural Plasticity

Article Title: Toll-Like Receptor 2 Attenuates Traumatic Brain Injury-Induced Neural Stem Cell Proliferation in Dentate Gyrus of Rats

doi: 10.1155/2020/9814978

Figure Lengend Snippet: Expression of TLR2 protein and mRNA in the DG (western blotting and PCR). (a) Western blotting: electrophoresis bands of TLR2 protein controlled with β -actin; (b) western blotting: the optical density of TLR2 electrophoresis; (c) real-time PCR: the expression of TLR2 mRNA and GAPDH was used as the endogenous reference gene. The TLR2 expression in the protein and mRNA level was significantly higher in the TBI group than that in the sham group ( ∗ p < 0.05), and the TLR2 expression was significantly different among various time points ( # p < 0.05). Data were expressed as mean ± SEM.

Article Snippet: Primary antibodies were used as follows: mouse anti-TLR2 antibody (1 : 500, Biorbyt, orb191498, San Francisco, CA, USA) and mouse anti- β -actin antibody (1 : 3000, TDY BIOTECH, TDY041, Beijing, China) overnight at 4°C.

Techniques: Expressing, Western Blot, Electrophoresis, Real-time Polymerase Chain Reaction

Journal: Neural Plasticity

Article Title: Toll-Like Receptor 2 Attenuates Traumatic Brain Injury-Induced Neural Stem Cell Proliferation in Dentate Gyrus of Rats

doi: 10.1155/2020/9814978

Figure Lengend Snippet: Gene sequences for primer synthesis.

Article Snippet: Primary antibodies were used as follows: mouse anti-TLR2 antibody (1 : 500, Biorbyt, orb191498, San Francisco, CA, USA) and mouse anti- β -actin antibody (1 : 3000, TDY BIOTECH, TDY041, Beijing, China) overnight at 4°C.

Techniques: Sequencing

Journal: International journal of molecular sciences

Article Title: Targeting TLR2/Rac1/cdc42/JNK Pathway to Reveal That Ruxolitinib Promotes Thrombocytopoiesis.

doi: 10.3390/ijms232416137

Figure Lengend Snippet: Figure 8. Biophysical validation reveals TLR2 as a new protein target of ruxolitinib. (A) Venn di‑ agram shows the common targets of Ruxolitinib and thrombocytopenia. The intersecting part rep‑ resents the common targets between Ruxolitinib and thrombocytopenia; (B) PPI network for identi‑ fying core targets of Ruxolitinib against thrombocytopenia through the screening conditions of De‑ gree > 47, BC > 0.002858932, CC > 0.507867733; (C) TLR2 and ligands (ruxolitinib) by molecular dock‑ ing; (D). Representative immunoblot images and biochemical quantification of TLR2 after treatment with Ruxolitinib (5, 10, and 20 µM) in Meg‑01 cells for 5 day (E) The DARTS assay for target valida‑ tion. TLR2 protein stability was increased upon Ruxolitinib (200 µM) treatment in Meg‑01 lysates. Pronase was added using several dilutions (1:500, 1:1000, or 1500) from 50 µg/mL stock for 10 min at 40 ◦C; (F) The DARTS assay demonstrated the dose‑dependent binding of Ruxolitinib to TLR2. Treatment with pronase (1:1000) was conducted for 10 min at 40 ◦C; (G) Meg‑01 cells were treated with ruxolitinib (20 µM), C29 (50 µM), ruxolitinib (20 µM) + C29 (50 µM) for 5 days. FCM analysis of the expression of CD41 and CD42b. (H) The histogram shows the percentage of CD41+/CD42b+

Article Snippet: The primary antibodies were as follows: FOS (Proteintech, USA, 66590‐1‐lg), EGR1 (Proteintech, USA, 22008–1‐AP), RUNX1 (Proteintech, USA, 25315‐1‐AP), TLR2 (Pro‐ teintech, USA, 17236‐1‐AP), Rac1/cdc42 (CST, USA, 4651S), JNK (Abmart, China, T55490), p‐JNK (Abmart, China, T55541), NF‐E2 (Proteintech, USA, 11089‐1‐AP), and GAPDH (Pro‐ teintech, USA, 60004‐1‐lg).

Techniques: Biomarker Discovery, Western Blot, Binding Assay, Expressing

Journal: International journal of molecular sciences

Article Title: Targeting TLR2/Rac1/cdc42/JNK Pathway to Reveal That Ruxolitinib Promotes Thrombocytopoiesis.

doi: 10.3390/ijms232416137

Figure Lengend Snippet: Figure 10. Schematic illustration of the role of ruxolitinib in MK differentiation and platelet pro‑duction. Ruxolitinib induces the expression of various cytokines and TLR2, activates the Rac1/cdc42/JNK signaling pathway, and leads to the expression of AP‑1, EGR1, RUNX1, and NF‑E2. As a result, the activation of AP‑1, EGR1, RUNX1, NF‑E2 promote the expression of genes related to MK differentiation and thrombopoiesis. These genes contribute to MK maturation and platelet for‑ mation and promote the recovery of bone marrow and spleen MKs and accelerate platelet production in RI‑mice. PPF: proplatelet‑forming MK.

Article Snippet: The primary antibodies were as follows: FOS (Proteintech, USA, 66590‐1‐lg), EGR1 (Proteintech, USA, 22008–1‐AP), RUNX1 (Proteintech, USA, 25315‐1‐AP), TLR2 (Pro‐ teintech, USA, 17236‐1‐AP), Rac1/cdc42 (CST, USA, 4651S), JNK (Abmart, China, T55490), p‐JNK (Abmart, China, T55541), NF‐E2 (Proteintech, USA, 11089‐1‐AP), and GAPDH (Pro‐ teintech, USA, 60004‐1‐lg).

Techniques: Expressing, Activation Assay

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