Journal: Molecular Pain

Article Title: Role of spinal cord alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in complete Freund's adjuvant-induced inflammatory pain

doi: 10.1186/1744-8069-4-67

Figure Lengend Snippet: The relative levels of GluR2 (A and B) and GluR1 (C and D) in the crude cytosolic (A and C) and plasma membrane fractions (B and D) from the ipsilateral L 4–5 dorsal horn at 2 and 24 h following saline or CFA injection . Naïve rats ( n = 4–8) were used as a control group. Data are presented as mean ± SEM. n = 4–8 rats/treatment. * p < 0.05, ** p < 0.01 vs the corresponding naïve group. E: The specificity of the fractionation procedure. Top: representative Western blots showing the expression of N -cadherin, PSD-95, and β-actin in total soluble fraction, plasma membrane fraction, and cytosolic fraction from L 4–5 dorsal horns of naïve rats ( n = 3). Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding protein levels in the total soluble fraction.

Article Snippet: The blotting membrane was blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-GluR1 (1:200; Upstate/CHEMICON, Temecula, CA), rabbit anti-GluR2 (1:500; Upstate/CHEMICON), rabbit anti- N -cadherin (1:1,000; BD Biosciences, Palo Alto, CA), mouse anti-PSD-95 (1:1,000; Upstate/CHEMICON), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Injection, Fractionation, Western Blot, Expressing

Journal: Cancers

Article Title: PD-L1 Expression with Epithelial Mesenchymal Transition of Circulating Tumor Cells Is Associated with Poor Survival in Curatively Resected Non-Small Cell Lung Cancer

doi: 10.3390/cancers11060806

Figure Lengend Snippet: CK pos /EpCAM pos /CD45 neg CTCs and CTC expression analysis for PD-L1, vimentin, and N-Cadherin in NSCLC patients. 7.5 mL blood was drawn, CTCs were enriched by microfilter isolation and immunofluorescence staining was performed for cytokeratins (CK) 8/18 and/or 19, EpCAM, CD45, and the nucleus identified with DAPI. Following identification of traditional CK pos /EpCAM pos /CD45 neg CTCs (left panels showing merged images), fluorescence quenching with borohydride, followed by re-staining by immunofluorescence for checkpoint inhibitor target PD-L1 and epithelial-mesenchymal transition (EMT) markers vimentin and N-Cadherin was performed. Different CTC expression patterns with regard to PD-L1, vimentin, and N-Cadherin are shown.

Article Snippet: Anti-PD-L1 antibody (rabbit mAb; clone 73-10; abcam, Cambridge, MA, USA), anti-vimentin (mouse mAb; clone V9; abcam), and anti-N Cadherin (mouse mAb; clone 5D5; abcam) were incubated at 4 °C overnight.

Techniques: Expressing, Isolation, Immunofluorescence, Staining, Fluorescence

Journal: Cancers

Article Title: PD-L1 Expression with Epithelial Mesenchymal Transition of Circulating Tumor Cells Is Associated with Poor Survival in Curatively Resected Non-Small Cell Lung Cancer

doi: 10.3390/cancers11060806

Figure Lengend Snippet: PD-L1 and EMT markers vimentin and N-Cadherin expressions determined by immunostaining in CTCs and patient-matched non-small cell lung cancer (NSCLC) tissues. Shown are representative images of expression patterns of immunohistochemically stained NSCLC tissues and patient-matched CTCs that were stained by immunofluorescence for PD-L1 and EMT markers.

Article Snippet: Anti-PD-L1 antibody (rabbit mAb; clone 73-10; abcam, Cambridge, MA, USA), anti-vimentin (mouse mAb; clone V9; abcam), and anti-N Cadherin (mouse mAb; clone 5D5; abcam) were incubated at 4 °C overnight.

Techniques: Immunostaining, Expressing, Staining, Immunofluorescence

Journal: Cancers

Article Title: PD-L1 Expression with Epithelial Mesenchymal Transition of Circulating Tumor Cells Is Associated with Poor Survival in Curatively Resected Non-Small Cell Lung Cancer

doi: 10.3390/cancers11060806

Figure Lengend Snippet: CTC counts, and comparative CTC expression and patient-matched tumor tissue analysis for PD-L1, vimentin, and N-Cadherin in NSCLC patients ( N = 30). ( A ) Counts per 7.5 mL of blood of traditional CK pos /EpCAM pos /CD45 neg CTCs are shown. CTC positive expression for PD-L1, vimentin, and N-Cadherin (defined as ≥50% mean intensity determined by quantification software) was determined after quenching of fluorescence and immunofluorescence re-staining with specific antibodies. PD-L1 pos CTCs were detected at a significantly higher rate than vimentin pos CTCs and/or N-Cadherin pos CTCs ( p -value was calculated by non-parametric Tukey’s multiple comparison analysis). EMT pos CTCs had to co-express both vimentin and N-Cadherin. PD-L1 pos /EMT pos CTCs were observed in 26/30 (86.7%) of NSCLC patients. No CTCs were identified in 15 healthy control subjects. ( B ) Comparative analysis of NSCLC tumor tissues and patient-matched CTCs ( N = 30) expression proportion (%) scores for PD-L1 (left panel), vimentin (middle panel), and N-Cadherin (right panel). PD-L1, vimentin, and N-Cadherin were statistically significantly higher expressed in CTCs than in patient-matched NSCLC tissues ( p -values were calculated with non-parametric Wilcoxon signed-rank test for matched pairs).

Article Snippet: Anti-PD-L1 antibody (rabbit mAb; clone 73-10; abcam, Cambridge, MA, USA), anti-vimentin (mouse mAb; clone V9; abcam), and anti-N Cadherin (mouse mAb; clone 5D5; abcam) were incubated at 4 °C overnight.

Techniques: Expressing, Software, Fluorescence, Immunofluorescence, Staining, Comparison, Control

Journal: Cancers

Article Title: PD-L1 Expression with Epithelial Mesenchymal Transition of Circulating Tumor Cells Is Associated with Poor Survival in Curatively Resected Non-Small Cell Lung Cancer

doi: 10.3390/cancers11060806

Figure Lengend Snippet: Expression proportion scores of patient-matched CTCs and NSCLC tumor tissues ( N = 30).

Article Snippet: Anti-PD-L1 antibody (rabbit mAb; clone 73-10; abcam, Cambridge, MA, USA), anti-vimentin (mouse mAb; clone V9; abcam), and anti-N Cadherin (mouse mAb; clone 5D5; abcam) were incubated at 4 °C overnight.

Techniques: Expressing