Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The expression of transporter associated with antigen processing 1 (TAP1) in pan-cancer. (A) TAP1 expression of 31 kinds of normal tissues downloaded from genotype-tissue expression (GTEx) database. (B) TAP1 expression in 21 kinds of cancer cell lines derived from the cancer cell line encyclopedia (CCLE). (C) TAP1 expression in tumor samples and the adjacent normal samples of 20 cancers derived from the TCGA. (D) TAP1 expression in tumor samples and normal samples of 27 cancers derived from the cancer genome atlas (TCGA) and GTEx database. * p < 0.05. ** p < 0.01. *** p < 0.001.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Expressing, Derivative Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The prediction ability of transporter associated with antigen processing 1 (TAP1) in pan-cancer prognosis. (A–C) The top three cancers correlated with TAP1 in disease-free survival (DFS). (D–F) The top three cancers correlated with TAP1 in disease-specific survival (DSS). (G–I) The top three cancers correlated with TAP1 in progression-free survival (PFS). (J–L) The top three cancers correlated with TAP1 in Overall survival (OS).

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The correlation of transporter associated with antigen processing 1 (TAP1) with immune infiltration in multiple cancers. The top three cancers with the strongest correlation with 6 immune infiltration cells which included B cells, CD4+T cells, CD8+T cells, neutrophils, macrophages, and dendritic cells. The cancers were colon adenocarcinoma (COAD), kidney renal clear cell carcinoma (KIRC), and breast invasive carcinoma (BRCA), respectively.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The relationship between TAP1 and ESTIMATE score in multiple cancers. The ESTIMATE score was used to determine the correlation of TAP1 with immune cells and stromal cells in 33 tumors. TAP1 was positively correlated with ESTIMATE scores in cervical squamous cell carcinoma (CESC), ovarian cancer (OC), uveal melanoma (UVM), testicular germ cell tumors (TGCT), and thyroid carcinoma (THCA) ( p < 0.05).

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The relationship between transporter associated with antigen processing 1 (TAP1) and immune checkpoint therapy in multiple cancers. (A) The relationship between TAP1 and 47 immune checkpoint genes. Color green represented negative correlation with TAP1 and color red represented positive correlation with TAP1. (B) The correlation of TAP1 with tumor mutation burden (TMB). (C) The correlation of TAP1 with microsatellite instability (MSI). * p < 0.05. ** p < 0.01. *** p < 0.001.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Mutagenesis

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: The relationship between transporter associated with antigen processing 1 (TAP1) and neoantigen in various cancers. TAP1 was positively related to neoantigens in ovarian cancer (OC), cervical squamous cell carcinoma (CESC), lung adenocarcinoma (LUAD), breast invasive carcinoma (BRCA), skin cutaneous melanoma (SKCM), and brain lower grade glioma (LGG) ( p < 0.05), whereas it was negatively correlated in colon adenocarcinoma (COAD) and rectum adenocarcinoma (READ).

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Immunopeptidomics

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: Correlation of transporter associated with antigen processing 1 (TAP1) expression with the prognosis of ovarian cancer (OC) and cervical cancer (CC). (A) Immunohistochemical (IHC) analysis of transporter associated with antigen processing 1 (TAP1) protein expression in OC tissues. Original magnifications:×200 and ×400. i represents strong positive staining, ii represents moderately positive staining, and iii represents weakly positive staining. (B) IHC analysis of TAP1 protein expression in CC tissues. Original magnifications:×200 and ×400. i represents strong positive staining, ii represents moderately positive staining, and iii represents weakly positive staining. Scale bars 200 μm. (C) 5-Year Kaplan–Meier analysis of overall survival in OC patients on the basis of TAP1 protein expression. (D) 5-Year Kaplan–Meier analysis of overall survival in CC patients on the basis of the TAP1 protein expression. (E) The protein-protein interaction (PPI) network of TAP1 and top 26 correlated transcription factors (TFs) were identified using STRING ( https://string-db.org/cgi/input.pl ). Color blue represented TFs negatively correlated with TAP1, color red represented TFs positively correlated with TAP1, and color white represented TFs significantly correlated with the top 26 TFs in enrichment analysis. (F) Gene set enrichment analysis of transporter associated with antigen processing 1 (TAP1) and pivotal TFs in OC. (G) TAP1, MEF2A, LEF1, and β-actin protein expression in normal cell lines and cancer cell lines. Error bars represent the SD of triplicate measurements. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: Downregulation of transporter associated with antigen processing 1 (TAP1) reduces the invasion and migration in ovarian cancer cells. (A) The volcano map of TAP1-related pathways by gene set variation analysis. (B) Enriching pathway analysis in ovarian cancer of TAP1. (C) The migration abilities of OVCAR3 and SKOV3 were photographed (i, ii) and measured (iii) by examining the wound closure after TAP1 knockdown using wound healing assays. Original magnifications, ×100. Scale bars 150 μm. (D) Transwell assays were photographed (i) and measured (ii) to detect the migration abilities after TAP1 knockdown in SKOV3 and OVCAR3 cells. Original magnifications, ×200. Scale bars 100 μm. (E) Colony formation assays were photographed (i) and measured (ii) to detect the proliferation abilities after TAP1 knockdown in SKOV3 and OVCAR3 cells. (F) Effects of TAP1 knockdown on migration-associated protein (E-cadherin and vimentin) and transcription factor (MEF2A and LEF1) were analyzed by western blotting in SKOV3 and OVCAR3 cells (i). Error bars of histogram (ii) represented the SD of triplicate measurements. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Migration, Knockdown, Western Blot

Journal: Frontiers in Molecular Biosciences

Article Title: Comprehensive Analysis of the Potential Immune-Related Biomarker Transporter Associated With Antigen Processing 1 That Inhibits Metastasis and Invasion of Ovarian Cancer Cells

doi: 10.3389/fmolb.2021.763958

Figure Lengend Snippet: To establish the model of tumor metastasis, SKOV3 cells were injected into nude mice by the i.p. route and randomly treated with si-TAP1_01 or si-NC ( n = 4/group). (A) Representative images of metastatic tumors in the peritoneal cavity. Yellow arrows and rectangles highlighted visible tumor nodes and liver metastasis. (B,C) Number of metastasis and total weight of all metastases were shown (mean ± SEM). (D) Body weight growth of all mice were shown (mean ± SEM). (E) The protein expression of migration-associated protein (E-cadherin and vimentin) and transcription factor (MEF2A and LEF1) in the tumor of si-TAP1_01 group and si-NC group (i). Error bars of histogram (ii) represented the SD of quadruplicate measurements. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The membrane was incubated with the following primary antibodies TAP1 (1:3,000), MEF2A (1:2000, #12382-1-AP, Proteintech, United States), LEF1 (1:1000, #14972-1-AP, Proteintech), E-cadherin (1:10000, #20874-1-AP, Proteintech), vimentin (1:2000, #10366-1-AP, Proteintech), and beta actin (1:1000, # 20536-1-AP, Proteintech).

Techniques: Injection, Expressing, Migration

Journal: Microbiology (Reading, England)

Article Title: Quantification of type VI secretion system activity in macrophages infected with Burkholderia cenocepacia.

doi: 10.1099/mic.0.000174

Figure Lengend Snippet: Fig. 4. Investigation of the B. cenocepacia VgrGs. (a) Measurement of T6SS activity. The infections were performed at an m.o.i. of 50 : 1 for 4 h with B. cenocepacia K56-2 DatsR and derivative vgrG mutants. The proportion of dark ‘beads on a string-like’ structures around macrophages was measured as described in Methods and in the legend of Fig. 1.The dotted line indicates the 0.2 relative unit threshold indicative of no ‘beads on a string-like’ structures. Values are means¡SD for at least 21 fields of view and are representative of three independent experiments. All the single vgrG deletion strains induced ‘beads on a string-like’ structures at a similar level to DatsR (no statistically significant differences) and significantly different levels (P,0.001) from uninfected cells. (b, c) Western blot analysis of total cell lysates (Pellet) and concentrated culture supernatants recovered from B. cenocepacia K56-2 DatsR, DatsRDhcp, vgrG mutants and the vgrG-less strain K56-2 DatsR D10vgrGs using anti-RNAP a subunit (cytosolic protein, cell lysis control) and anti-Hcp antibodies.

Article Snippet: After blocking (Roche), the membranes were incubated with the following primary antibodies as required: 4RA2 monoclonal antibody (Neoclone), which cross-reacts with the B. cenocepacia and B. multivorans RNA polymerase subunit alpha (cytosolic/cell lysis control; dilution 1 : 25 000); anti-Hcp polyclonal antiserum (dilution 1 : 1 000; ProSci) and FLAG M2 monoclonal antibody (dilution 1 : 50 000; Sigma).

Techniques: Activity Assay, Western Blot, Lysis, Control

Journal:

Article Title: Calcium signalling through nucleotide receptor P2X1 in rat portal vein myocytes

doi: 10.1111/j.1469-7793.2001.0339c.xd

Figure Lengend Snippet: A, amplified DNA fragments of P2X receptors (lanes 1–7) from rat brain (a) and rat portal vein myocytes (b) were separated on a 2 % agarose gel and visualized by staining with ethidium bromide. Lane 8, RNA from brain and portal vein myocytes in the absence of reverse transcriptase served as a negative control. Numbers on the left indicate molecular size standards in base pairs (bp). For RNA purification and PCR conditions, see Methods. B, immunostaining of P2X receptor subtypes in portal vein myocytes. Myocytes were stained with anti-P2X1 receptor (a) or anti-P2X4 receptor antibody (b) and vizualization was realized with a donkey anti-rabbit IgG FITC-conjugated antibody. In the absence of primary antibodies or after inactivation of the antibodies by their antigen peptides, only a faint background fluorescence was observed (not shown). Typical confocal sections were performed above the nucleus and therefore appeared spherical. Both P2X1 (a) and P2X4 (b) receptor subtypes were distributed throughout the confocal sections with a marked staining of P2X1 receptor subtype at the cell periphery.

Article Snippet: The rabbit anti-P2X1 and anti-P2X4 receptor antibodies (Alomone Labs, Jerusalem, Israel) were directed against polypeptides corresponding to residues 382–399 and 370–388 of the rat P2X1 and P2X4 receptors, respectively.

Techniques: Amplification, Agarose Gel Electrophoresis, Staining, Negative Control, Purification, Immunostaining, Fluorescence

Journal: The Journal of Neuroscience

Article Title: Presynaptic L-Type Ca 2+ Channels Increase Glutamate Release Probability and Excitatory Strength in the Hippocampus during Chronic Neuroinflammation

doi: 10.1523/JNEUROSCI.2981-19.2020

Figure Lengend Snippet: Long-lasting LPS treatment increases the expression of L-type Ca2+ channels at glutamatergic synapses. A, Top, Representative images of neurites of hippocampal neurons exposed to either vehicle or LPS for 48 h and double-immunostained with Cav1.2 antibodies (green) and the excitatory presynaptic marker vGLUT1 (red). Merge panels represent synaptic puncta in which Cav1.2 and vGLUT1 colocalize (yellow; arrows). Bottom, Representative immunoreactivity intensity profiles of the colocalization experiments showing the increased occurrence of overlap between Cav1.2- and vGLUT1-positive puncta in the samples chronically treated with LPS. B, Quantification of the density of vGLUT1-positive puncta counted on 30 μm dendrite tracts starting from the neuronal body. Data are mean ± SEM from three independent experiments, each conducted in duplicate (veh = 30; LPS = 29). C, Quantification of the absolute Cav1.2/vGLUT1 colocalization area (top) and of the Cav1.2/vGLUT1 colocalization expressed in percent of the total Cav1.2-immunopositive area (bottom) in vehicle- and LPS-treated neurons (veh = 66; LPS = 97). Data are mean ± SEM. **p < 0.01 (unpaired Student's t test/Mann–Whitney U test).

Article Snippet: The presynaptic localization of the L-type Ca 2+ channel was analyzed in treated neurons by evaluating the immunoreactivity of L-type Ca 2+ channel (1:200, Alomone Labs) ( Hermosilla et al., 2017 ) in vGLUT1-positive excitatory boutons.

Techniques: Expressing, Marker, MANN-WHITNEY

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: A , lung static elastance (Est); B , viscoelastic component of elastance (ΔE); C - E , resistive (ΔP1), viscoelastic/inhomogeneous (ΔP2), and total lung pressures (ΔPtot), respectively in wild-type (WT) and P2X7 receptor knockout (KO) animals. Values represent mean + SEM of 6–9 animals/group (10 determinations per animal). *p <0.05 and **p <0.001 in relation to the respective control (CTRL).

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Representative lung parenchyma photomicrographs (hematoxylin-eosin staining) of: A and B , wild-type (WT) and P2X7 receptor knockout (KO) mice after saline instillation, respectively; C-E , WT mice after silica instillation showing polymorphonuclear (arrows), mononuclear cells (arrowhead), and areas of cellular debris (asterisk); D-F , P2X7 receptor KO mice after silica instillation showing polymorphonuclear cells (arrowhead), mononuclear cells (arrows), and focus of apoptotic cells (asterisk). Bars: 1300 µm ( A and B ); 1700 µm ( C and D ); 330 µm ( E and F ). G - H , nodular score and free silica particles quantification, respectively, in lung parenchyma of both genotypes. Box plots represent median of 6 to 9 animals per group (15 pictures/animal) with a statistical cut of ± 10-90% demonstrating their respective SD. * p <0.05.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Staining, Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma (PicroSirius) of wild-type ( A, B ) and P2X7 receptor knockout ( C, D ) 14 days after intratracheal instillation of silica particles ( B, D ) or saline ( A, C ). E : quantification of collagen fibers in lung parenchyma of wild-type (WT) and P2X7 receptor knockout (KO) mice. Box plots show median values of 5–7 animals in each group with statistical cut off of ±10–90%, with respective SD (15 random non-coincident fields/animal), and are expressed as % surface density of fibers per tissue area in each field. Bars: 900 µm; *p <0.05 and **p<0.01 in relation to the respective control; ++ p <0.05 in relation to Silica-WT.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma of wild-type mice 14 days after intratracheal instillation of saline (CTRL, A ) or silica particles (SiO 2 , B, C ). D and E : negative control of immunohistochemistry reaction and primary antibody + peptide (1∶1), respectively. Bars: 950 µm ( A, B ); 750 µm ( C, D and E ). F: quantification of P2X7 receptor immunoreactivity. Box plots represent median of 6 to 9 animals per experimental group with a statistical cut of ± 10-90%, with respective SD (15 random non-coincident fields/animal). * p <0.05.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Negative Control, Immunohistochemistry

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma of wild-type (WT; A, C, E ) and P2X7 receptor knockout (KO; B, D, F ) mice 14 days after intratracheal instillation of silica particles ( C-F ) or saline ( A, B ). Arrows: mononuclear cells. Asterisk: aggregates of reactive cells. Bars: 1250 µm ( A, B ); 600 µm ( C, D ); 90 µm ( E, F ). G: quantification of iNOS immunoreactivity. Box plots represent median of 6-9 animals per experimental group with a statistical cut of ± 10-90%, with respective SD (15 random non-coincident fields/animal). **p <0.01 and ***p <0.001 in relation to the respective control; ++ p <0.01 in relation to Silica WT.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma of wild-type (WT; A, C and E ) and P2X7 receptor knockout (KO; B, E and F ) mice 14 days after instillation of silica particles ( C-F ) or saline ( A, B ). Bars: 1100 µm ( A, B ); 180 µm ( C-F ). Arrows: reactive inflammatory ( C, E, F ) and epithelial cells ( D ). Asterisk: aggregates of reactive cells. G: quantification of p-Smad2/3 immunoreactivity in silica WT and KO mice. Box plots show median values of 5–7 animals in each group with a statistical cut off of ±10–90%, with respective SD (15 random non-coincident fields/animal). ***p <0.001.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma of wild-type ( A, C ) and P2X7 receptor knockout ( B, D ) mice 14 days after intratracheal instillation of silica particles. Immunoreactivity was present mainly in mononuclear (arrow) and polymorphonuclear (arrowhead) cells in nodular area ( A, B -inserts). Asterisks show reactive inflammatory cells in WT ( C ) and KO mice ( D ). E, F : reactive bronchiolar (arrow) and smooth muscle (asterisk) cells in WT and KO mice, respectively. Bars: 700 µm ( A ); 650 µm ( B ); 30 µm ( A, B -inserts); 350 µm ( C and D ); 120 µm ( E and F ). G: quantification of NF-κB immunoreactivity in lung parenchyma. Box plots show median of 5–7 animals in each group with a statistical cut off of ±10–90%, with respective SD (15 random non-coincident fields/animal). **p <0.01 and ***p<0.001 in relation to the respective control; ++ p <0.01 in relation to Silica WT.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Photomicrographs of lung parenchyma of wild-type ( A, B, E ) and P2X7 receptor knockout ( C, D ) mice 14 days after intratracheal instillation of silica particles. Arrows: TUNEL positive cells. E : reactive brochiolar cells (arrow) in WT mice. Bars: 1250 µm ( A ); 130 µm ( B ); 200 µm ( C, D ); 70 µm ( E ). F : quantification of apoptosis (TUNEL immunoreactivity) in lung parenchyma. Values are mean + SEM of 6-9 animals per group (15 random non-coincident fields/animal). *p <0.05.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out, TUNEL Assay

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Values are mean + SEM of 5–7 mice in wild-type (WT) and P2X7 receptor knockout (KO) groups 14 days after intratracheal instillation of saline (CTRL) or silica particles (SILICA). *p <0.05 in relation to the respective control; #p <0.05 in relation to Silica WT.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Quantification of IL-1β in supernatant of alveolar macrophages ( A ), and fibroblasts ( B ) treated with silica (SiO 2 ) or ATP, in the presence or not of the P2X7 receptor antagonist A740003. Values are mean + SEM of four independent experiments.*p <0.05; #p <0.05 in relation to no treatment.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques:

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Quantification of NO secretion in supernatant from alveolar macrophages: ( A ) directly treated with silica particles (SiO 2 ) or ATP, in the presence or not of P2X7 receptor inhibitor A740003; or ( C ) treated with supernatant obtained from silica-treated macrophages. Flow cytometry quantification of: B , ROS production (measured by mean fluorescence intensity, MFI); and D , percentage of hypodiploid cell formation (apoptosis) in alveolar macrophage culture after different treatments. oATP: periodate oxidized ATP (P2X7 receptor inhibitor). Values are mean + SEM of three independent experiments. *p <0.05 in relation to no treatment; #p <0.05 in relation to the same agonist without oATP.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Flow Cytometry, Fluorescence

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Macrophages were exposed to silica particles and imaged using differential interference contrast (DIC). (A-C) Light micrographs of macrophages from wild-type (WT), and (D-E) P2X7 receptor knockout (KO) mice exposed to saline (CTRL) or silica particles (SIL) in the absence or presence of the P2X7 receptor inhibitor oATP (SIL+oATP). Note increased vesicle formation in silica treated macrophages from WT mice (B, arrow) in comparison with macrophages from P2X7 KO mice (E, arrow). B, insert: macrophage pre-treated with cytochalasin and exposed to silica. Bars: 20 µm.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Knock-Out

Journal: PLoS ONE

Article Title: P2X7 Receptor Modulates Inflammatory and Functional Pulmonary Changes Induced by Silica

doi: 10.1371/journal.pone.0110185

Figure Lengend Snippet: Silica-particles induce ATP release. Extracellular ATP activates P2X7 receptors, culminating in P2X7-mediated ROS production, inflammasome activation, and IL-1β release.

Article Snippet: Four-micrometer-thick sections were collected onto poly-l-lysine prepared slides and stained with specific P2X7 receptor (Alomone, Jerusalem, Israel), nitric oxide synthase inducible (iNOS) (Thermo Fisher Scientific Inc., AL, USA), phospho-Smad2/3 (p-Smad2/3, nuclear protein that activates transforming growth factor-β, TGF-β) (R&D Systems Inc, Minneapolis, MN, USA), and nuclear transcription factor-κB (NF-κB) antibodies (Thermo Fisher Scientific Inc., AL, USA).

Techniques: Activation Assay

Journal: Science translational medicine

Article Title: The anti-CD40L monoclonal antibody AT-1501 promotes islet and kidney allograft survival and function in nonhuman primates

doi: 10.1126/scitranslmed.adf6376

Figure Lengend Snippet: (A) CD40L binding potency of AT-1501 (cyan) and Hu5C8 (orange) was determined by ELISA with recombinant human CD40L (rhuCD40L). Abatacept (purple), a variant of CTLA4-Ig that interferes with the CD28-CD80/86 costimulatory pathway, was used as a negative control. A450, absorbance at 450nm. (B) AT-1501 and Hu5C8 binding to human FcγRI, FcγRIIa, FcγRIIIa, and FcγRIIIb was measured by ELISA. (C to F) PAC-1 expression was measured in platelets from healthy controls incubated with soluble CD40L alone (C), Hu5C8 and sCD40L (D), Hu5C8 F(Ab’)2 lacking an Fc region and sCD40L (E), or AT-1501 and sCD40L (F). Blue tracings represent expression of PAC-1 following incubation with 5μg/mL sCD40L alone and red tracings represent expression of PAC-1 following incubation with the indicated combinations. (G) Platelet aggregation was measured in vitro using platelets incubated with the indicated combinations.

Article Snippet: ELISAs were run with Hu5C8 (ProSci, XP-5263BT), an anti-CD40L antibody, as a positive control.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Variant Assay, Negative Control, Expressing, Incubation, In Vitro