Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 1 | Leukocyte recruitment after anti-MPO antibody administration. (a) Glomerular neutrophil recruitment to C57BL/6 wild-type (WT, n ¼ 6) mice. Highly purified lipopolysaccharide (hpLPS) and anti-OVA (aOVA) antibodies (n ¼ 6) or anti-MPO (aMPO) antibodies alone (n ¼ 6) increased glomerular neutrophil recruitment compared with untreated WT animals (n ¼ 4), represented as a dotted line (Po0.001). Co-administration of hpLPS and aMPO antibodies (n ¼ 6) induced more neutrophil recruitment compared with hpLPS and aOVA antibodies or aMPO antibodies alone. (b) Pulmonary MPO activity in WT mice. Untreated WT mice had 0.62 U of MPO activity/g (dotted line); MPO activity was increased in all antibody injected groups, compared with untreated mice. Significantly more pulmonary MPO activity was seen in the WT mice treated with hpLPS and aMPO antibodies compared with other treatment groups. Photomicrographs representative of glomerular neutrophil recruitment, with one glomerular neutrophil in WT mice treated with hpLPS and aOVA antibodies (c) or aMPO antibodies alone (d), or three neutrophils in mice treated with LPS and aMPO antibodies (e) are demonstrated. In panels f and g glomerular and lung neutrophil recruitment in WT mice are compared with recruitment in TLR4/ mice (n ¼ 4). Neutrophil recruitment in untreated in TLR4/ mice (n ¼ 5) did not differ from untreated WT mice (dotted line). Glomerular neutrophil recruitment (f) and lung MPO activity (g) decreased in TLR4/ mice compared with WT mice. *Po0.05, ***Po0.001. Original magnification x400. gcs, glomerular cross-section; MPO, myeloperoxidase; OVA, ovalbumin; TLR, Toll-like receptor.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Purification, Activity Assay, Injection

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 2 | TLR4 staining and expression in the kidney. Positive TLR4 staining (green) was detectable in glomeruli of WT mice treated with LPS and anti-MPO antibodies (a). No TLR4 staining was visible in TLR4/ mice treated with anti-MPO antibodies (b). Glomeruli from WT mice were stained with anti-CD31 antibodies to identify endothelial cells (red staining) (c). Merged image of TLR4 and endothelial cell staining that identifies endothelial cell TLR4 production (yellow) (d). After stimulation with LPS and anti-MPO (aMPO) antibodies there was an increase in TLR4 mRNA expression in microdissected murine glomeruli, and little change in TLR4 expression was seen in the tubulointerstitium (n ¼ 4) (e). In human conditionally immortalized ciGEnC lines, baseline TLR4 expression was increased compared with both podocytes and mesangial cells (which did not express TLR4), n ¼ 6 for all experimental groups (f). After LPS stimulation TLR4 expression increased only in endothelial cells. (g) TLR4 protein expression by ciGEnC. Flow cytometric analysis of cultured ciGEnC incubated with no antibody (i), isotype control antibody (ii), and anti-human TLR4 antibody (iii) demonstrated TLR4 protein expression. *Po0.05, **Po0.01, ***Po0.001. Original magnification x800. Ab, antibody; ciGEnC, conditionally immortalized glomerular endothelial cell, LPS, lipopolysaccharide; MFI, mean florescence intensity; MPO, myeloperoxidase; TLR, Toll-like receptor; WT, wild type.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Staining, Expressing, Cell Culture, Incubation, Control

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 3 | CXCL1 and CXCL2 kidney mRNA expression and immunostaining. Using tissues from experiments detailed in Figure 1, kidney CXCL1 mRNA expression was increased in WT mice after hpLPS and anti-OVA (aOVA) antibodies (Po0.001), n ¼ 6, aMPO antibodies alone (Po0.05), n ¼ 6, and hpLPS and aMPO antibodies (Po0.001), n ¼ 6. Compared with WT mice given hpLPS and aMPO antibodies, TLR4/

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Expressing, Immunostaining

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 4 | Glomerular endothelial cell, CXCL1, CXCL2, and TLR4 colocalization. Kidneys from WT mice given LPS and anti-MPO antibodies were stained for (a) CD31 (blue), (b) TLR4 (green), and (c) CXCL1 (red). Endothelial cells and CXCL1 colocalized (magenta) (d). CXCL1 and TLR4 also colocalized (yellow) (e), and a merged three-color image showed that some endothelial cells were positive for both CXCL1 and TLR4 (white) (f). To assess CXCL2 production kidneys were stained for (g) CD31 (blue), (h) TLR4 (green), and (i) CXCL2 (red). CXCL2 colocalized with glomerular endothelial cells (magenta) (j) and TLR4 (yellow) (k). Merged three-color image showing that some endothelial cells were positive for both CXCL2 and TLR4 (white) (l). Original magnification x800. LPS, lipopolysaccharide; MPO, myeloperoxidase; TLR, Toll-like receptor; WT, wild type.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Staining

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 6 | Assessment of glomerular leukocyte recruitment, lung MPO activity, and glomerular chemokine staining in BM chimeric wild type (WT)-WT mice (bone marrow (BM) þ , tissue cell (TC) þ , n ¼ 8), TLR4/-WT mice (BM–TC þ , n ¼ 8), and WT-TLR4/ mice (BM þ TC–, n ¼ 8) injected with hpLPS and anti-MPO (aMPO) antibodies. Glomerular neutrophil recruitment (a) was decreased in both BM–TC þ and BM þ TC– compared with BM þ TC þ mice. There was decreased glomerular neutrophil recruitment in BM–TC þ mice compared with BM þ TC– mice. Lung MPO activity was decreased in both BM–TC þ and BM þ TC– mice compared with BM þ TC þ mice (b). Kidney CXCL1 (c) and CXCL2 (d) immunostaining was decreased in BM–TC þ and BM þ TC– mice compared with BM þ TC þ mice. *Po0.05, **Po0.01, ***Po0.001. gcs, glomerular cross-section; MPO, myeloperoxidase.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Activity Assay, Staining, Injection, Immunostaining

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 10 | Functional and histological renal injury in BM chimeric mice wild type (WT)-WT mice (bone marrow (BM) þ , tissue cell (TC) þ , n ¼ 7), Toll-like receptor (TLR4/) -WT mice (BM–TC þ , n ¼ 6), and WT-TLR4/ mice (BM þ TC–, n ¼ 7) injected with hpLPS and aMPO antibodies. Compared with ‘sham chimeras’ (BM þ TC þ ) 24 h albuminuria (a) was decreased in BM–TC þ and BM þ TC– chimeras. There was a non-significant trend to decrease in dipstick hematuria (b). Glomerular fibrin deposition (c) and glomerular hypercellularity (d) was decreased in BM–TC þ and BM–TC þ chimeric mice compared with sham chimeras BM þ TC þ . *Po0.05, **Po0.01, ***Po0.001.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Functional Assay, Injection

Journal: Kidney international

Article Title: Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis.

doi: 10.1038/ki.2010.327

Figure Lengend Snippet: Figure 8 | Functional and histological renal injury in WT and TLR4/ mice treated with LPS and anti-MPO antibodies. At 6 days after the administration of aMPO antibody/LPS 24 h albuminuria (a), dipstick hematuria (b), glomerular fibrin deposition (c), and glomerular hypercellularity (d) was attenuated in TLR4/ mice (n ¼ 6) compared with WT controls (n ¼ 6). The dotted line in (a) and (d) represents mean values in untreated WT mice. Representative figures are shown demonstrating glomerular injury in WT mice (e) compared with TLR4/ mice (f). More glomerular fibrin deposition was seen in WT mice (g) compared with TLR4/ mice (h). *Po0.05, **Po0.01, ***Po0.001. Original magnification x400. gcs, glomerular cross-section; LPS, lipopolysaccharide; MPO, myeloperoxidase; TLR, Toll-like receptor; WT, wild type.

Article Snippet: For CXCL1 colocalization, antibodies were rabbit anti-mouse TLR4 (20 mg/ml), and goat anti-mouse CXCL1 (20 mg/ml, R&D Systems), then chicken anti-rabbit Alexa Fluor 488 and chicken anti-goat Alexa Fluor 594 (1:200, Molecular Probes).

Techniques: Functional Assay

Journal: PloS one

Article Title: Stiffening-induced high pulsatility flow activates endothelial inflammation via a TLR2/NF-κB pathway.

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: Figure 2. Proximal stiffening induces proinflammatory response and upregulates TLR2 expression in downstream bovine PAECs. (A-B) High pulsatility flow (HPF), due to the use of a “stiff” tube upstream to cell culture, upregulated proinflammatory molecule mRNA (ICAM-1, VCAM-1, MCP-1 and E-selectin) and protein (MCP-1) in healthy PAECs, compared to low pulsatility flow (LPF) or to static conditions. (C-D) The mRNA and protein expression of TLR2 but not TLR4 in PAECs was highly upregulated by HPF. *: p<0.05 versus Static, †: p<0.05 versus LPF. https://doi.org/10.1371/journal.pone.0102195.g002

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Expressing, Cell Culture

Journal: PloS one

Article Title: Stiffening-induced high pulsatility flow activates endothelial inflammation via a TLR2/NF-κB pathway.

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: Figure 3. Pharmacological or siRNA inhibition of TLR2 results in suppression of PAEC proinflammatory responses caused by stiffening-induced HPF. (A, C) At the mRNA level, TLR2/4 inhibitor OxPAPC or TLR2 siRNA but not TLR4 inhibitor CLI-095, decreased PAEC expression of ICAM-1, VCAM-1, MCP-1 and E-selectin mRNAs under HPF; TLR4 siRNA decreased ICAM-1 and E-selectin but not VCAM-1 and MCP-1 mRNAs. “*”: p<0.05 versus LPF, “†”: p<0.05 versus HPF. (B, D) At the protein level, the MCP-1 expression in PAECs exposed to HPF was inhibited by OxPAPC or TLR2 siRNA treatment but not CLI-095 or TLR4 siRNA. The black line in the blot images (D, right) shows separated lanes obtained on the same gel. https://doi.org/10.1371/journal.pone.0102195.g003

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Inhibition, Expressing

Journal: PloS one

Article Title: Stiffening-induced high pulsatility flow activates endothelial inflammation via a TLR2/NF-κB pathway.

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: Figure 6. Enhanced TLR and MCP-1 expression in the distal pulmonary artery endothelium in vivo. (A) PAECs from calves with hypoxia-induced pulmonary hypertension (PH) show elevated expression of TLR2 and TLR4, compared to control (CO). *:p<0.05. (B) Both immunostaining and western blotting results show elevated MCP-1 expression by PH-ECs compared to CO-ECs from calves. “PA” indicates the lumen of a pulmonary artery. *:p<0.05. (C) Enhanced TLR2 expression in the pulmonary arterial endothelium of human with pulmonary arterial hypertension (PAH). Cryosections of human intra-lobar pulmonary arteries were immunostained with TLR2 (red fluorescence) and counterstained with DAPI (cell nuclei, blue). Elastic lamellae showed green auto-fluorescence. https://doi.org/10.1371/journal.pone.0102195.g006

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Expressing, In Vivo, Control, Immunostaining, Western Blot

Journal: bioRxiv

Article Title: Urokinase receptor associates with TLR4 interactome to promote LPS response

doi: 10.1101/2020.06.10.143826

Figure Lengend Snippet: A. Primary peritoneal macrophages from uPAR−/− mice were stimulated with suPAR with or without LPS for 15 min. Then, cells were fixed and stained for TLR4 (Alexa 488, green) and uPAR (Alexa 647, red). DAPI was used as nuclear stain. Scale bar 10 μm. B. Primary WT and uPAR−/− macrophages were stimulated with 100 ng/ml LPS and1μg/ml suPAR for 3 hrs C. Raw 264.7 cells were stimulated with 100 ng/ml LPS, fixed and stained as in A. Scale bar 12.5 μm. D. Raw 264.7 were stimulated with 1μg/ml biotin-LPS for 30 min, then cell lysis was performed. Protein complexes were precipitated using Streptavidin magnetic beads and analyzed by western blotting using anti-murine-uPAR antibody.

Article Snippet: Unconjugated and Alexa 647-conjugated mouse uPAR antibody were from R&D systems (MAB531 and FAB531R, respectively); TLR4 antibody (MAB27591) was from SrnD Systems; LPS (L2887) was from Sigma, Biotin-LPS and PAM3CSK4 were from Invivogen; Soluble mouse uPAR was from CinoBiologicals.

Techniques: Staining, Lysis, Magnetic Beads, Western Blot

Journal: bioRxiv

Article Title: Urokinase receptor associates with TLR4 interactome to promote LPS response

doi: 10.1101/2020.06.10.143826

Figure Lengend Snippet: A. Biotin-LPS binding was assessed in SiCo and uPARsi HK-2 cells as described. B. Duolink proximity ligation assay to assess uPAR/TLR4 and uPAR/CD36 interaction was performed on HK-2 cells stimulated with LPS for 15 min as described in Methods. C. Duolink images were quantified using Particles analysis tool of ImageJ. D. HK-2 cells were stimulated with LPS for 3 hrs after cell pre-treatment with CD36 inhibitor SSO. Expression of IL-6 and IL-8 was assessed by TaqMan RT-PCR.

Article Snippet: Unconjugated and Alexa 647-conjugated mouse uPAR antibody were from R&D systems (MAB531 and FAB531R, respectively); TLR4 antibody (MAB27591) was from SrnD Systems; LPS (L2887) was from Sigma, Biotin-LPS and PAM3CSK4 were from Invivogen; Soluble mouse uPAR was from CinoBiologicals.

Techniques: Binding Assay, Proximity Ligation Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Urokinase receptor associates with TLR4 interactome to promote LPS response

doi: 10.1101/2020.06.10.143826

Figure Lengend Snippet: A. Peritoneum of sham and LPS-injected WT mice was fixed and stained for uPAR and TLR4. DAPI used as nuclear stain­ scale bar 100μm. B. Expression of TNFα, MCP-1, IL-6 and IL-1 0 was assessed in mouse blood plasma before and 20 h after CLP surgery using Cytometric Beads Array. C: IL-6/IL-10 ratio in CLP mice 20 hrs after surgery.

Article Snippet: Unconjugated and Alexa 647-conjugated mouse uPAR antibody were from R&D systems (MAB531 and FAB531R, respectively); TLR4 antibody (MAB27591) was from SrnD Systems; LPS (L2887) was from Sigma, Biotin-LPS and PAM3CSK4 were from Invivogen; Soluble mouse uPAR was from CinoBiologicals.

Techniques: Injection, Staining, Expressing, Clinical Proteomics

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: Oligonucleotide primers used for the quantitative real time PCR.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques:

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 transcript are expressed in endometrium during the menstrual cycle . Columns indicate mean TLR3 and TLR4 mRNA quantities from endometrium in proliferative (n = 16), secretory (n = 11) and menstrual phase (n = 8) run in triplicates. The y-axis is scaled logarithmically; error bars represent the standard deviation of the mean.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 protein is localised to endometrial cells during the menstrual cycle . TLR3 protein staining in healthy late proliferative (LP) tissue was high in luminal and glandular tissue (A , brown precipitate) and lower in LP endometriotic tissue (B) . Late secretory (LS) endometrium showed highly expressed TLR3 in the epithelium (C) , but weakly in endometriosis (D) . Intense staining of TLR4 proteins was shown in mid proliferative (MP) tissue (E) . In late proliferative phase of endometriosis, TLR4 proteins were comparably lower (F) . TLR4 protein was high in mid secretory (MS) normal endometrium (G) , whereas it was decreased in endometriotic MS tissue (H) . During the menstrual phase, both TLR3 (I) and TLR4 (J) were highly expressed. Co-immunostaining for TLR4 (green), CD14 ( K , red) and CD163 ( L , red) demonstrated that TLR4 proteins were expressed by CD14 positive dendritic cells and monocytes ( K , yellow) and by CD163 positive macrophages ( L , yellow). Localisation of TLR4 to immune cells is marked by a black arrow (J) and by white arrows (K, L) .

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining, Immunostaining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 mRNA expression is regulated in endometriosis . The expression of TLR3 (A, C) and TLR4 mRNA (B, D) in endometrium during proliferative (n = 13, run in triplicates, A, B) and secretory phase (n = 3, C, D) was decreased in eutopic endometriotic endometrium when compared to controls. In addition, four proliferative corresponding lesions were evaluated (A, B) showing a local upregulation of both receptors on ectopic sites. Columns represent the mean ratio of TLR copy number to ACTB copy number. Error bars represent the standard deviation of the mean. * P < 0.05; ** P < 0.01.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Expressing, Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 protein is locally induced in endometriotic lesions . No specific TLR3 protein staining was seen in eutopic endometriotic tissue (A) whereas a high glandular localisation of the protein was detected in a gland of an ectopic endometriotic lesion from the same patient (B) . Similarly, TLR4 was not detectable in eutopic endometrium (C) but present in glandular epithelium of ectopic endometrium from the same women (D) .

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 mRNA expression is decreased in endometrial adenocarcinoma . (A-B) Columns indicate mean TLR3 (A) and TLR4 (B) mRNA levels from postmenopausal patients (PMP, n = 8), and those diagnosed with endometrial hyperplasia (HP, n = 10) and endometrial carcinoma (EnCa, n = 16). (C-D) TLR3 (C) and TLR4 (D) mRNA expression in different carcinoma grades compared to postmenopausal controls and hyperplastic endometrium: G1 (n = 5), G2 (n = 6) and G3 (n = 5). Error bars represent the standard deviation of the mean. * P < 0.05; ** P < 0.01.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Expressing, Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 proteins are present in postmenopausal endometrium . Localisation of TLR3 in normal postmenopausal endometrium (A) , endometrial hyperplasia (B) , endometrial adenocarcinoma grade G1 (C) , G2 (D) and G3 (E ). Localisation of TLR4 protein in normal postmenopausal endometrium (F) , endometrial hyperplasia (G) , endometrial adenocarcinoma grade G1 (H) , G2 (I) and G3( J ). All stained sections indicated epithelium as the preferred localisation of TLR3 and TLR4 proteins. TLR4 protein was additionally present in immune cells (arrows).

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR4 is localised to immune cells of postmenopausal endometrium . Co-Immunostaining of TLR4 with CD14 and CD163 in healthy endometrium (A, B) and in adenocarcinoma (C, D) . TLR4 proteins were expressed by CD14 positive dendritic cells and monocytes (A, C) and by CD163 positive macrophages (B, D) . Arrows indicate the co-localisation of TLR4 with the immune cells. Scale bar = 20 μm.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Immunostaining

Journal: bioRxiv

Article Title: A novel reporter gene assay for pyrogen detection

doi: 10.1101/613190

Figure Lengend Snippet: Expression of TLR2 (green line), TLR4 (orange line), and TLR6 (purple line) on RAW 264.7 cells (a) and binding curves of the corresponding antibodies (Ab, concentrations from 10 μg/ml to 0.005 μg/ml, at 1:3 dilutions) to TLR2, TLR4, and TLR6 on RAW 264.7 cells (b) were analysed by FACS. Rat IgG2a AF488 (blue shadow or line) was used as an isotype control.

Article Snippet: Then, 50 μl of 2 μg/ml fluorescence-labelled antibodies was added for the detection of TLR2 (R&D, Cat # FAB1530G), TLR4 (R&D, Cat # FAB27591G), and TLR6 (R&D, Cat # FAB1533G); rat IgG2a Alexa Fluor (AF) 488-conjugated antibody was used as an isotype control (R&D, Cat # IC006G).

Techniques: Expressing, Binding Assay