Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Clinicopathological features according to TLR4 and MyD88 expression

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Detection of TLR4 and MyD88 immunoreactivity in normal colon tissues, adenoma, and CRC. ( A ) Normal colon mucosa (left) and adenoma (right) showed no or weak immunoreactions for TLR4 (top) and MyD88 (below), which was identified as negative expression in this study. Original magnification, × 200. ( B ) In contrast to the situation in the normal and adenoma, strong immunostaining of TLR4, which was localized in the membrane (top) or cytoplasm (below), was observed in the cancer cells. Original magnification, right, × 200; left, × 400. ( C ) Strong immunostaining of MyD88, which was localised in the cytoplasm, was observed in the cancer cells. Original magnification, right, × 200; left, × 400. ( D ) High expression of TLR4 was correlated with high expression of MyD88 in CRC. CRC with a high expression of TLR4 ( n =22) showed significantly high levels of MyD88 expression than did CRC with a low expression of TLR4 ( n =86) ( P <0.05). CRC with a high expression of MyD88 ( n =25) showed significantly high levels of TLR4 expression than did CRC with a low expression of MyD88 ( n =83) ( P <0.005).

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques: Expressing, Immunostaining, Membrane

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Differences in immunostaining scores of TLR4 and MyD88 in liver metastasis (+) CRC ( n =22) and liver metastasis (−) CRC ( n =86) were analysed. The TLR4 ( A ), MyD88 ( B ), and combined TLR4/MyD88 expression ( C ) levels were significantly higher in liver metastasis (+) CRC than in liver metastasis (−) CRC ( P =0.0015, P =0.0035, P =0.0001, respectively).

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques: Immunostaining, Expressing

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Clinicopathological features, tumour markers, and patient survival (univariate analysis)

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques:

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Clinicopathological features, tumour markers, and patient survival (multivariate analysis)

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques:

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Kaplan–Meier survival curves of DFS and OS in patients with CRC according to TLR4 and MyD88 expression. ( A , B ) High expression of TLR4 was potentially associated with poor DFS and significantly associated with poor OS ( P =0.013). ( C , D ) High expression of MyD88 was significantly associated with poor DFS and OS ( P =0.0029, P =0.0001, respectively). ( E , F ) High co-expression of TLR4+MyD88 was significantly associated with poor DFS and OS ( P =0.0042, P =0.0001, respectively).

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

doi: 10.1038/sj.bjc.6605558

Figure Lengend Snippet: Clinicopathological features, tumour markers, and patient survival (multivariate analysis)

Article Snippet: The tissue sections were then incubated with a serum-free protein blocker and incubated at 4°C overnight with mouse monoclonal anti-human TLR4 antibody (1:100; HTA125, eBioscience, San Diego, CA, USA) or rabbit polyclonal anti-MyD88 antibody (1:100; HFL-296, Santa Cruz Biotech, Santa Cruz, CA, USA).

Techniques:

Journal: Physiological Reports

Article Title: IL‐33/ST2 receptor‐dependent signaling in the development of pulmonary hypertension in Sugen/hypoxia mice

doi: 10.14814/phy2.15185

Figure Lengend Snippet: Pulmonary hypertension parameters in SUHX mice. Developed right ventricular pressures in (a) C57Bl/6J, (b) ST2 −/− , and (c) MYD88 −/− mice in response to SUHX and DMSO/RA. Right ventricle contractility [(RV) + dP/dt] in response to SUHX and DMSO/RA treatments in (d) C57Bl/6J, (e) ST2 −/− , and (f) MYD88 −/− mice. Right ventricle relaxation [(RV) − dP/dt] in response to SUHX and DMSO/RA in (g) C57Bl/6J, (h) ST2 −/− , and (i) MYD88 −/− mice. Values are presented as mean ± SD ( n = 15 male, n = 8 female DMSO/RA; n = 14 male, n = 6 female SUHX). *Significant difference between SUHX and DMSO/RA within a sex ( p < 0.05)

Article Snippet: Left lung lobe extracts were prepared in RIPA Buffer with Protease Inhibitor (Sigma‐Aldrich), electrophoresed by SDS‐PAGE with Tris Carboxy Ethyl Phosphene (TCEP; Thermo Fisher Scientific), electrotransferred to a Fluorescence PVDF membrane (MilliporeSigma), blocked for 60 min (Blocking Buffer; LI‐COR Biosciences) and probed with the following antibodies: IL‐33 (mouse mAb, 1:200 dilution, Nessy‐1 ALX‐804‐840/1; Enzo Life Sciences, Inc.), ST2 (goat pAb, 1:1000, Human ST2/IL‐33R AF523; R&D Systems), MYD88 (goat pAb, 1:1000, Mouse/Rat MyD88 AF3109; R&D Systems) and α ‐tubulin (rabbit Ab, 1:2000 dilution; Cell Signaling Technology) overnight at 4°C.

Techniques:

Journal: Physiological Reports

Article Title: IL‐33/ST2 receptor‐dependent signaling in the development of pulmonary hypertension in Sugen/hypoxia mice

doi: 10.14814/phy2.15185

Figure Lengend Snippet: Pulmonary vascular remodeling‐wall thickness in the small arteries (<50 μm in diameter) is shown in lung sections stained with hematoxylin and eosin (a) C57Bl/6J, (b) ST2 −/− and (c) MYD88 −/− mice under DMSO/RA and SUHX treatments. Quantitation of wall thickness in (d) C57Bl/6J, (e) ST2 −/− and (f) MYD88 −/− under DMSO/RA and SUHX conditions. Values are presented as mean ± SD ( n = 9–14 DMSO; n = 14–17 SUHX). **Statistical significance between DMSO/RA and SUHX treatments within each genotype, p < 0.05

Article Snippet: Left lung lobe extracts were prepared in RIPA Buffer with Protease Inhibitor (Sigma‐Aldrich), electrophoresed by SDS‐PAGE with Tris Carboxy Ethyl Phosphene (TCEP; Thermo Fisher Scientific), electrotransferred to a Fluorescence PVDF membrane (MilliporeSigma), blocked for 60 min (Blocking Buffer; LI‐COR Biosciences) and probed with the following antibodies: IL‐33 (mouse mAb, 1:200 dilution, Nessy‐1 ALX‐804‐840/1; Enzo Life Sciences, Inc.), ST2 (goat pAb, 1:1000, Human ST2/IL‐33R AF523; R&D Systems), MYD88 (goat pAb, 1:1000, Mouse/Rat MyD88 AF3109; R&D Systems) and α ‐tubulin (rabbit Ab, 1:2000 dilution; Cell Signaling Technology) overnight at 4°C.

Techniques: Staining, Quantitation Assay

Journal: Physiological Reports

Article Title: IL‐33/ST2 receptor‐dependent signaling in the development of pulmonary hypertension in Sugen/hypoxia mice

doi: 10.14814/phy2.15185

Figure Lengend Snippet: Vascular remodeling in small arteries and arterioles. Endothelial cell proliferation was evaluated by immunohistochemical and flow cytometry analysis of proliferating CD31 + cells. The percentage of proliferating endothelial cells (% CD31 + /BrDU + ) in (a) C57Bl/6J and (b) ST2 −/− mice under DMSO/RA and SUHX conditions. Localization of proliferating (Ki‐67) cells detected in SMA positive small arteries of (c) C57Bl/6J, (d) ST2 −/− , and (e) MYD88 −/− mice under DMSO/RA and SUHX conditions. Scale bars represent 50 μm. Values are presented as the means ± SD ( n = 5–15 DMSO/RA, n = 12–17 SUHX). **Statistical significance between DMSO/RA and SUHX treatments, p < 0.01

Article Snippet: Left lung lobe extracts were prepared in RIPA Buffer with Protease Inhibitor (Sigma‐Aldrich), electrophoresed by SDS‐PAGE with Tris Carboxy Ethyl Phosphene (TCEP; Thermo Fisher Scientific), electrotransferred to a Fluorescence PVDF membrane (MilliporeSigma), blocked for 60 min (Blocking Buffer; LI‐COR Biosciences) and probed with the following antibodies: IL‐33 (mouse mAb, 1:200 dilution, Nessy‐1 ALX‐804‐840/1; Enzo Life Sciences, Inc.), ST2 (goat pAb, 1:1000, Human ST2/IL‐33R AF523; R&D Systems), MYD88 (goat pAb, 1:1000, Mouse/Rat MyD88 AF3109; R&D Systems) and α ‐tubulin (rabbit Ab, 1:2000 dilution; Cell Signaling Technology) overnight at 4°C.

Techniques: Immunohistochemical staining, Flow Cytometry

Journal: Physiological Reports

Article Title: IL‐33/ST2 receptor‐dependent signaling in the development of pulmonary hypertension in Sugen/hypoxia mice

doi: 10.14814/phy2.15185

Figure Lengend Snippet: IL‐33 protein levels in whole lungs of mice exposed to SUHX (a) Western Blot representation of IL‐33, ST2, and MYD88 protein in lungs of C57BL/6J, ST2 −/− , and MYD88 −/− mice exposed to either SUHX or DMSO/RA conditions (all n = 8). Equal loading of protein was confirmed using α‐Tubulin. (b) Quantitative analysis of IL‐33 in whole lungs. Values are presented as the means ± SD

Article Snippet: Left lung lobe extracts were prepared in RIPA Buffer with Protease Inhibitor (Sigma‐Aldrich), electrophoresed by SDS‐PAGE with Tris Carboxy Ethyl Phosphene (TCEP; Thermo Fisher Scientific), electrotransferred to a Fluorescence PVDF membrane (MilliporeSigma), blocked for 60 min (Blocking Buffer; LI‐COR Biosciences) and probed with the following antibodies: IL‐33 (mouse mAb, 1:200 dilution, Nessy‐1 ALX‐804‐840/1; Enzo Life Sciences, Inc.), ST2 (goat pAb, 1:1000, Human ST2/IL‐33R AF523; R&D Systems), MYD88 (goat pAb, 1:1000, Mouse/Rat MyD88 AF3109; R&D Systems) and α ‐tubulin (rabbit Ab, 1:2000 dilution; Cell Signaling Technology) overnight at 4°C.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Oleacein Attenuates Lipopolysaccharide-Induced Inflammation in THP-1-Derived Macrophages by the Inhibition of TLR4/MyD88/NF-κB Pathway †

doi: 10.3390/ijms23031206

Figure Lengend Snippet: Modulation of LPS-induced activation of MyD88 and NF-κB in THP-1-derived macrophages by OLC. THP-1-derived macrophages were treated with different concentrations of OLC (1, 2.5 and 5 μM) for 30 min prior LPS (100 ng/mL) for 24 h. The results from MyD88 measured by ELISA assay ( A ) and real-time PCR ( B ) are shown along with results of DNA binding activity of NF-κB p65 ( C ) obtained by ELISA tests. Data are the mean ± SEM of three independent experiments performed in triplicate. *** p < 0.001 vs. ctrl; § p < 0.05 vs. LPS-treated cells; §§ p < 0.01 vs. LPS-treated cells; §§§ p < 0.001 vs. LPS-treated cells.

Article Snippet: Lysates were then centrifuged at 10,000× g for 5 min, and the supernatants were collected and analyzed for quantification of Myd88 through human MyD88 ELISA Kit (ab171341; Abcam, Cambridge, UK).

Techniques: Activation Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Binding Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Oleacein Attenuates Lipopolysaccharide-Induced Inflammation in THP-1-Derived Macrophages by the Inhibition of TLR4/MyD88/NF-κB Pathway †

doi: 10.3390/ijms23031206

Figure Lengend Snippet: Oligonucleotide primer sequences used for the real-time PCR.

Article Snippet: Lysates were then centrifuged at 10,000× g for 5 min, and the supernatants were collected and analyzed for quantification of Myd88 through human MyD88 ELISA Kit (ab171341; Abcam, Cambridge, UK).

Techniques: Sequencing