Journal: Nature Communications

Article Title: Synapse type-specific proteomic dissection identifies IgSF8 as a hippocampal CA3 microcircuit organizer

doi: 10.1038/s41467-020-18956-x

Figure Lengend Snippet:

Article Snippet: Primary antibodies were the following: mouse anti-PSD95 (1:2500; Thermo-Fisher), rabbit ant-Synaptophysin (1:2500; Sigma-Aldrich), rabbit anti-Synpr (1:2500; Synaptic Systems), rabbit anti-Nectin3 (1:500; Abcam), goat anti-MBP (1:500; Santa Cruz), rabbit anti-Histone H3 (1:1000; Cell Signaling Technology), goat anti-IgSF8 (1:2000; R&D Systems), mouse anti-TenR (1:1000; R&D Systems), rabbit anti-BRINP2 (1:100; Atlas Antibodies), sheep anti-CNTN1 (1:500; R&D Systems), goat anti-ICAM5 (1:500; R&D Systems), sheep anti-ISLR2 (1:1000; R&D Systems), goat anti-Kit/SCFR (1:1000; R&D Systems), goat anti-NEGR1 (1:1000; R&D Systems), goat anti-Neogenin (1:1000; R&D Systems), goat anti-NRP1 (1:500; R&D Systems), goat anti-PlexinA1 (1:500; R&D Systems), rabbit anti-PTPRD (1:500; Novus), goat anti-PTPRS (1:1000; R&D Systems), rabbit anti-ROBO2 (1:1000; Abcam), and sheep anti-Teneurin-4 (1:1000; R&D Systems).

Techniques: Virus, Plasmid Preparation, Recombinant, Transfection, Antibody Labeling, Cloning, Suspension, Cell Culture, Knock-Out, Software, Microscopy

Journal: Cancers

Article Title: UCHL5 Promotes Proliferation and Migration of Bladder Cancer Cells by Activating c-Myc via AKT/mTOR Signaling.

doi: 10.3390/cancers14225538

Figure Lengend Snippet: Figure 4. Knockdown of UCHL5 results in inactivation of the AKT/mTOR signaling pathway and contributes to decreased c-Myc, SLC25A19, and ICAM5 expression. (A) KEGG pathway enrichment analysis was used to assess RNA-Seq data containing shControl and shUCHL5 bladder cancer cells. (B,C) GSEA identified enrichment of AKT/mTOR signaling pathway and c-Myc gene sets. (D) Heat map of the differential expressed genes. (E) The difference in AKT/mTOR signaling pathway scores was defined using ssGSEA between the UCHL5 high- and UCHL5 low-expression groups. (F) The association of UCHL5 expression and AKT/mTOR signaling pathway score. (G) qRT-PCR determination of the mRNA levels of c-Myc, SLC25A19, and ICAM5 in bladder cancer cells transfected with shControl or shUCHL5. (H–K) Protein levels of the AKT/mTOR signal pathway proteins (t-PI3K, p-PI3K, t-AKT, p-AKT, t-mTOR, and p-mTOR), c-Myc, SLC25A19, and ICAM5 in UCHL5 silenced and overexpressed bladder cancer cells shown by Western blotting. β-Actin is used as the loading control. Values are shown as mean ± SD. * p < 0.05 and **** p < 0.0001 compared with controls.

Article Snippet: Antibodies against β-actin (Cat. AC038), UCHL5 (Cat. A7978), SLC25A29 (Cat. A12373), ICAM5 (Cat. A4156), PI3K (Cat. A4860), p-PI3K (Cat. AP0854), AKT (Cat. 17909), p-AKT (S473, Cat. AP0637), mTOR (Cat. A11355), and P-mTOR (Cat. AP0115) were acquired from ABclonal (Abclonal, United States).

Techniques: Knockdown, Expressing, RNA Sequencing, Quantitative RT-PCR, Transfection, Western Blot, Control

Journal: Cancers

Article Title: UCHL5 Promotes Proliferation and Migration of Bladder Cancer Cells by Activating c-Myc via AKT/mTOR Signaling.

doi: 10.3390/cancers14225538

Figure Lengend Snippet: Figure 5. SC79 recovers the effect of UCHL5 knockdown in bladder cancer cells. (A) Transwell assay (Scale bar = 200 µm) assessment of cell migration in shCtrl, shUCHL5-3, and shUCHL5-3 + SC79 groups. (B,C) Cell proliferation of three groups evaluated using CCK-8 and colony formation assays. (D) Western blotting analysis of the p-AKT, t-AKT, p-mTOR, t-mTOR, c-Myc, SLC25A19, and ICAM5 protein expression in bladder cancer cells. β-Actin is used as the loading control. Values are shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with controls.

Article Snippet: Antibodies against β-actin (Cat. AC038), UCHL5 (Cat. A7978), SLC25A29 (Cat. A12373), ICAM5 (Cat. A4156), PI3K (Cat. A4860), p-PI3K (Cat. AP0854), AKT (Cat. 17909), p-AKT (S473, Cat. AP0637), mTOR (Cat. A11355), and P-mTOR (Cat. AP0115) were acquired from ABclonal (Abclonal, United States).

Techniques: Knockdown, Transwell Assay, Migration, CCK-8 Assay, Western Blot, Expressing, Control

Journal: Cancers

Article Title: UCHL5 Promotes Proliferation and Migration of Bladder Cancer Cells by Activating c-Myc via AKT/mTOR Signaling.

doi: 10.3390/cancers14225538

Figure Lengend Snippet: Figure 6. LY294002 rescues the effect of UCHL5 upregulation in bladder cancer cells. (A) Transwell as- say was used to assess cell migration of Vector, UCHL5, and UCHL5 + LY294002 (Scale bar = 200 µm). (B,C) Cell proliferation of three groups evaluated using CCK-8 and colony formation assays. (D) Western blotting analysis of the UCHL5, p-AKT, t-AKT, p-mTOR, t-mTOR, c-Myc, SLC25A19, and ICAM5 protein expression in bladder cancer cells. β-Actin is used as the loading control. Values are shown as mean ± SD. * p < 0.05, ** p < 0.01, and **** p < 0.0001 compared with controls.

Article Snippet: Antibodies against β-actin (Cat. AC038), UCHL5 (Cat. A7978), SLC25A29 (Cat. A12373), ICAM5 (Cat. A4156), PI3K (Cat. A4860), p-PI3K (Cat. AP0854), AKT (Cat. 17909), p-AKT (S473, Cat. AP0637), mTOR (Cat. A11355), and P-mTOR (Cat. AP0115) were acquired from ABclonal (Abclonal, United States).

Techniques: Migration, Plasmid Preparation, CCK-8 Assay, Western Blot, Expressing, Control

Journal: Frontiers in Neurology

Article Title: The TARC/sICAM5 Ratio in Patient Plasma is a Candidate Biomarker for Drug Resistant Epilepsy

doi: 10.3389/fneur.2012.00181

Figure Lengend Snippet: Assay of sICAM5 in plasma from epilepsy patients and controls . (A) Dot-plot of sICAM5 concentrations in plasma (square, patients; diamond, controls). For values between the horizontal lines, the assay does not accurately discriminate between patients and controls. (B) Bar graph and error calculation for data in Part (A) . Difference is significant for p = 0.002. (C) ROC curve for data in Part a, showing an area under the curve (AUC) value of 0.87.

Article Snippet: Wells were coated overnight with protein G affinity purified mouse monoclonal anti-human sICAM5 antibody (capture antibody; R&D Systems, Minneapolis, MN, USA; catalog # MAB 1950), 2 μg/mL diluted in phosphate buffered saline (PBS; 25 μL/well).

Techniques: Clinical Proteomics

Journal: Frontiers in Neurology

Article Title: The TARC/sICAM5 Ratio in Patient Plasma is a Candidate Biomarker for Drug Resistant Epilepsy

doi: 10.3389/fneur.2012.00181

Figure Lengend Snippet: Levels of cytokines and chemokines in plasma that significantly distinguish between epilepsy patients and controls .

Article Snippet: Wells were coated overnight with protein G affinity purified mouse monoclonal anti-human sICAM5 antibody (capture antibody; R&D Systems, Minneapolis, MN, USA; catalog # MAB 1950), 2 μg/mL diluted in phosphate buffered saline (PBS; 25 μL/well).

Techniques: Clinical Proteomics, Concentration Assay

Journal: Frontiers in Neurology

Article Title: The TARC/sICAM5 Ratio in Patient Plasma is a Candidate Biomarker for Drug Resistant Epilepsy

doi: 10.3389/fneur.2012.00181

Figure Lengend Snippet: IL-6/sICAM5 ratio in plasma from epilepsy patients and controls . (A) Dot-plot of IL-6/sICAM5 concentrations in plasma (square, patients; diamond, controls). For values between the horizontal lines, the assay does not accurately discriminate between patients and controls. (B) Bar graph and error calculation for data in Part (A) . Difference is not significant, with p = 0.087. (C) ROC curve for data in Part (A) , showing an area under the curve (AUC) value of 0.90.

Article Snippet: Wells were coated overnight with protein G affinity purified mouse monoclonal anti-human sICAM5 antibody (capture antibody; R&D Systems, Minneapolis, MN, USA; catalog # MAB 1950), 2 μg/mL diluted in phosphate buffered saline (PBS; 25 μL/well).

Techniques: Clinical Proteomics

Journal: Frontiers in Neurology

Article Title: The TARC/sICAM5 Ratio in Patient Plasma is a Candidate Biomarker for Drug Resistant Epilepsy

doi: 10.3389/fneur.2012.00181

Figure Lengend Snippet: IL-8/sICAM5 ratio in plasma from epilepsy patients and controls . (A) Dot-plot of IL-8/sICAM5 concentrations in plasma (square, patients; diamond, controls). For values between the horizontal lines, the assay does not accurately discriminate between patients and controls. (B) Bar graph and error calculation for data in Part (A) . Difference not significant, with p = 0.125. (C) ROC curve for data in Part (A) , showing an area under the curve (AUC) value of 0.88.

Article Snippet: Wells were coated overnight with protein G affinity purified mouse monoclonal anti-human sICAM5 antibody (capture antibody; R&D Systems, Minneapolis, MN, USA; catalog # MAB 1950), 2 μg/mL diluted in phosphate buffered saline (PBS; 25 μL/well).

Techniques: Clinical Proteomics

Journal: Frontiers in Neurology

Article Title: The TARC/sICAM5 Ratio in Patient Plasma is a Candidate Biomarker for Drug Resistant Epilepsy

doi: 10.3389/fneur.2012.00181

Figure Lengend Snippet: TARC/sICAM5 ratio in plasma from epilepsy patients and controls . (A) Dot-plot of TARC/sICAM5 concentrations in plasma (square, patients; diamond, controls). There are no values between the horizontal lines, indicating the assay accurately discriminates between patients and controls. (B) Bar graph and error calculation for data in Part (A) . Difference is 13-fold, and is significant for p = 0.034. (C) ROC curve for data in Part (A) , showing an area under the curve (AUC) value of 1.00.

Article Snippet: Wells were coated overnight with protein G affinity purified mouse monoclonal anti-human sICAM5 antibody (capture antibody; R&D Systems, Minneapolis, MN, USA; catalog # MAB 1950), 2 μg/mL diluted in phosphate buffered saline (PBS; 25 μL/well).

Techniques: Clinical Proteomics