elisa Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher elisa plate reader
    Recognition of wt rAAV2, rAAV8, and their chimeras by <t>ADK8.</t> (A) ADK8 <t>ELISA</t> for rAAV8 and rAAV8 with AAV2 amino acid substitution mutants. Wild-type rAAV8 and the mutants rAAV8 456GTANTQ461→GTTTQS and rAAV8 493TTTGQ497→KTSAD were detected by the ADK8, but the rAAV8 586LQQQNT591→LQRGNR mutant was not. (B) ADK8 ELISA for rAAV2, rAAV8, and rAAV2 with AAV8 amino acid substitution mutants. Wild-type rAAV2 and the rAAV2 457QSRLQ461→TQTLG mutant were not recognized by ADK8, but the rAAV2 585RGNRQ589→QQNTA mutant was. (C) ADK8 ELISA for rAAV8 and seven-residue peptide insertion (at aa 590) mutants. All insertions disrupted AAV8 detection by ADK8. In panels A to C, the mean standard deviations from four independent experiments are shown; asterisks identify mutants for which capsids could not be detected in the ADK8 ELISA (detection limit, 10 8 capsids/ml). A total of 10 10 viral genomes were used for each ELISA.
    Elisa Plate Reader, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2377 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa plate reader/product/Thermo Fisher
    Average 99 stars, based on 2377 article reviews
    Price from $9.99 to $1999.99
    elisa plate reader - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    90
    Millipore elisa
    Recognition of wt rAAV2, rAAV8, and their chimeras by <t>ADK8.</t> (A) ADK8 <t>ELISA</t> for rAAV8 and rAAV8 with AAV2 amino acid substitution mutants. Wild-type rAAV8 and the mutants rAAV8 456GTANTQ461→GTTTQS and rAAV8 493TTTGQ497→KTSAD were detected by the ADK8, but the rAAV8 586LQQQNT591→LQRGNR mutant was not. (B) ADK8 ELISA for rAAV2, rAAV8, and rAAV2 with AAV8 amino acid substitution mutants. Wild-type rAAV2 and the rAAV2 457QSRLQ461→TQTLG mutant were not recognized by ADK8, but the rAAV2 585RGNRQ589→QQNTA mutant was. (C) ADK8 ELISA for rAAV8 and seven-residue peptide insertion (at aa 590) mutants. All insertions disrupted AAV8 detection by ADK8. In panels A to C, the mean standard deviations from four independent experiments are shown; asterisks identify mutants for which capsids could not be detected in the ADK8 ELISA (detection limit, 10 8 capsids/ml). A total of 10 10 viral genomes were used for each ELISA.
    Elisa, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 4778 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa/product/Millipore
    Average 90 stars, based on 4778 article reviews
    Price from $9.99 to $1999.99
    elisa - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    93
    Thermo Fisher enzyme linked immunosorbent assays
    Efferocytosis of apoptotic mesenchymal stromal cells polarizes monocytes into an immunosuppressive phenotype. (A) Apoptotic mesenchymal stromal cells (ApoMSC) were stained with 20 ng/mL pHrodo™ Red-succinimidyl ester (PE) while isolated monocytes were stained with 1 μM CellTrace™ Violet (V450). Monocytes were cultured with different amounts of ApoMSC for 2 h at 37°C, 5% CO2. Lipopolysaccharide (LPS) 100 ng/mL was used to stimulate monocytes. The mean fluorescence intensity (MFI) (PE) within the monocyte population was measured as an indicator of efferocytosis, n=3. (B) MFI (PE) after 2 and 24 h was compared among the different groups, n=3. (C) Confocal images of monocytes efferocytosing ApoMSC after 24 h in culture. The yellow arrows indicate the monocytes engulfing ApoMSC while the white arrows indicate ApoMSC alone. The bar represents 20 μm. (D) Monocytes (2 × 10 5 /well) were co-cultured with ApoMSC (4 × 10 5 /well) for 8 h. CellTrace™ Violet-labeled CD3 T cells (2 × 10 5 /well) were added to the cultures and stimulated by CD3/CD28 beads at a 1:1 cell:bead ratio for 3 days. The percentages of proliferating T cells were measured by flow cytometry according to the loss of fluorescence intensity due to cell division, n=4. (E-G) Intracellular and surface staining was performed to examine the expression of different proteins, including COX2, IDO and PD-L1, in monocytes. The COX2 high , n=3 (E), IDO high , n=4 (F) and PD-L1 high , n=3 (G) populations were gated according to the monocyte alone control (used as a negative control). (H-J) Enzyme-linked <t>immunosorbent</t> assays were performed to assess the amount of PGE2, n=4 (H), IL-10, n=3 (I) and TNF-α, n=6 (J) in cell culture supernatants. (K) To select the monocytes according to efferocytosis, ApoMSC were fluorescent-labeled with CellTrace™ FarRed (APC + ) while monocytes were labeled with CellTrace™ Violet (V450 + ). APC + monocytes were regarded as efferocytosis + and APC − monocytes were regarded as efferocytosis − . (L) Bar charts showing the difference of COX2 high , IDO high and PD-L1 high populations between the efferocytosis + and efferocytosis- monocytes, n=3. Experimental data are expressed as means ± standard deviation. An unpaired t test was used to compare the mean differences between two samples. One-way analysis of variance and the post-hoc Tukey test were used to compare the mean differences among the samples (* P
    Enzyme Linked Immunosorbent Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1389 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enzyme linked immunosorbent assays/product/Thermo Fisher
    Average 93 stars, based on 1389 article reviews
    Price from $9.99 to $1999.99
    enzyme linked immunosorbent assays - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    95
    Thermo Fisher elisa kits
    Antibody responses, virus titers, and inflammatory cytokine levels in BALF and lungs after challenge. Levels of IgG antibodies were determined in samples collected from mice at day 5 p.i. with A/Philippines/2/82 (H3N2) virus ( n = 5). (A) IgG antibody levels in BALF. The antibody level was determined by <t>ELISA</t> using M2e peptide as a coating antigen. Lung viral titers (B), IFN-γ (C), and IL-6 (D) <t>cytokines</t> in BALF were determined at day 5 p.i. Lung viral titers were determined by an egg infection assay. IFN-γ and IL-6 were determined by a cytokine ELISA. Data represent mean ± SEM. Statistical significances were determined by 1-way ANOVA. Asterisks indicate significant differences (* p
    Elisa Kits, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 10950 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kits/product/Thermo Fisher
    Average 95 stars, based on 10950 article reviews
    Price from $9.99 to $1999.99
    elisa kits - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    92
    Becton Dickinson enzyme linked immunosorbent assays
    Expression of macrophage-inflammatory protein-2 (MIP-2) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. MIP-2 enzyme-linked <t>immunosorbent</t> assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P
    Enzyme Linked Immunosorbent Assays, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enzyme linked immunosorbent assays/product/Becton Dickinson
    Average 92 stars, based on 96 article reviews
    Price from $9.99 to $1999.99
    enzyme linked immunosorbent assays - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    94
    BioTek Instruments elisa reader
    Effects of EA on inflammatory cytokines levels in the peri-infarct hippocampal CA1and sensorimotor cortex of MCAO/R rats. a Representative and quantitative analysis of <t>ELISA</t> showed that EA treatment for 7 days decreased the levels of <t>IL-1β</t> and b increased the levels of IL-10 in MCAO/R injured rats. Data are presented as mean ± S.E.M. from 6 individual rats in each group. P
    Elisa Reader, supplied by BioTek Instruments, used in various techniques. Bioz Stars score: 94/100, based on 6839 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa reader/product/BioTek Instruments
    Average 94 stars, based on 6839 article reviews
    Price from $9.99 to $1999.99
    elisa reader - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    92
    BioTek Instruments elisa plate reader
    Antigenic analysis of Fwt and Flys-GCN. F protein preparations Fwt and Flys-GCN were coated in 96 well plates for <t>ELISA</t> analysis. The reactivity of the recombinant proteins with the different antibodies (indicated above each graph) was determined by applying 2-fold dilutions of these antibodies as indicated in the Materials and Methods. Binding of the antibodies was detected using <t>HRP-conjugated</t> secondary antibodies. The antibody dilution is indicated on the X-axes, while the Y-axes corresponds to optical density read at 450nm (OD450 nm). Error bars indicate standard deviations (n = 3).
    Elisa Plate Reader, supplied by BioTek Instruments, used in various techniques. Bioz Stars score: 92/100, based on 4836 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa plate reader/product/BioTek Instruments
    Average 92 stars, based on 4836 article reviews
    Price from $9.99 to $1999.99
    elisa plate reader - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Bio-Rad elisa reader
    miR-146a, miR-335 and miR-622 regulate THP-1 cell migration, invasion and proliferation. (A–C) THP-1 cells were transiently transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Then, cells were loaded in Boyden chamber on uncoated filters (A and B) or Matrigel coated filters (C). Then, cells were allowed to migrate towards the uPA aminoterminal fragment (ATF) or the stroma derived factor 1 (SDF1) (A) or towards 10% serum (FBS) (B and C). Migrated cells were fixed, stained with haematoxylin, and counted. Results of migration assays are expressed as percentage of cells migrated towards chemoattractants over the cells migrated without chemoattractants; 100% values represent cell migration in the absence of chemoattractants. The values are the mean ± SEM of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test. (D) THP-1 cells were transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Cells, harvested at indicated times, were loaded in 96-well plates and incubated with 20 μl/well of CellTiter 96 AQueous One Solution Reagent for 4 hrs at <t>37°C,</t> 5% CO 2 . Then, the absorbance was determined by an <t>ELISA</t> reader (Bio-Rad) at a wavelength of 490 nm. Fold increase represents the ratio between the OD at indicated time points and OD at time 0. The values are the mean of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test.
    Elisa Reader, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 4535 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa reader/product/Bio-Rad
    Average 92 stars, based on 4535 article reviews
    Price from $9.99 to $1999.99
    elisa reader - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    94
    Becton Dickinson elisa kits
    Effects of ASWP and three ASEPs on cell viability; phagocytic activity; and the production of nitric oxide (NO), tumor necrosis factor <t>(TNF)-α,</t> interleukin (IL)-6, and IL-12 in RAW264.7 macrophages. ASWP was obtained from Aster scaber using hot water extraction and ASEP-A, ASEP-C, and ASEP-P were obtained from Aster scaber using α-amylase, cellulase, and pectinase, respectively, in an enzyme-assisted extraction. RAW264.7 macrophages were stimulated with the polysaccharide samples at 1, 10, or 100 μg/mL. LPS (1 μg/mL) was used as the positive control (PC). ( A ) Cell viability was determined by the cell counting kit-8 (CCK-8) assay; ( B ) Phagocytic activity was determined using a phagocytosis assay; ( C ) NO levels in the culture media were determined by measuring nitrite accumulation, and the secretion levels of TNF-α, IL-6, and IL-12 were measured by <t>ELISA.</t> ( D ) mRNA expression levels of iNOS, TNF-α, IL-6, and IL-12 in macrophages were quantified by qRT-PCR analysis. Lowercase letters (a–c) indicate significant differences ( p
    Elisa Kits, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 3147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kits/product/Becton Dickinson
    Average 94 stars, based on 3147 article reviews
    Price from $9.99 to $1999.99
    elisa kits - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    94
    Bio-Rad elisa plate reader
    Effect of curcumin on A549 cell viability. The cytotoxic effects of curcumin on A549 cells were examined by <t>MTT</t> assay. Cells were cultured in growth medium at different concentrations (0–80 μM) for 12 h and 24 h, and absorbance at 540 nm was measured using an <t>ELISA</t> reader. Bar graphs indicate the percentage of viability. All data are expressed as mean ± SEM of three independent experiments. *** p
    Elisa Plate Reader, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 3289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa plate reader/product/Bio-Rad
    Average 94 stars, based on 3289 article reviews
    Price from $9.99 to $1999.99
    elisa plate reader - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    95
    Abcam elisa kit
    Detection of secreted albumin and blood urea nitrogen by <t>ELISA.</t> The <t>ALB</t> and BUN concentration of UC-MSCs and BM-MSCs culture supernatants collected on weeks 1, 2, 3, 4 and 5 following hepatic differentiation were measured. The medium of hepatocyte was measured as a control. Each bar represents the mean ± standard deviation (n=3). *P
    Elisa Kit, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 1611 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kit/product/Abcam
    Average 95 stars, based on 1611 article reviews
    Price from $9.99 to $1999.99
    elisa kit - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    92
    Abcam enzyme linked immunosorbent assays
    The synthesized GILZ-p decreased LPS induced inflammatory cytokine secretion in culture medium of Müller cells. The Enzyme-Linked <t>Immunosorbent</t> Assays (ELISA) were performed to determine the protein expression levels of IL-1β, MCP-1, TNF-α, and ICAM-1 in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 6 for each group. ∗ P
    Enzyme Linked Immunosorbent Assays, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enzyme linked immunosorbent assays/product/Abcam
    Average 92 stars, based on 45 article reviews
    Price from $9.99 to $1999.99
    enzyme linked immunosorbent assays - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    Millipore elisa kit
    Effect of Imm124-E on serum levels of IL-6, <t>GLP-1,</t> and adiponectin. All sera were measured using <t>ELISA</t> kits at days 1 and 30 of the trial in all treated patients. Graphs show ( A ) Serum levels of IL-6 from six patients; ( B ) Serum levels of GLP-1 post–glucose tolerance test from five patients; ( C ) Serum levels of adiponectin from eight patients. Note: Graphs indicate means ± SD. Abbreviations: IL-6, interleukin-6; GLP-1, glucagon-like peptide 1; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation.
    Elisa Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2068 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kit/product/Millipore
    Average 99 stars, based on 2068 article reviews
    Price from $9.99 to $1999.99
    elisa kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    94
    Cayman Chemical elisa kit
    Acetylation and expression of StAR in response to HDAC inhibitors, and their correlation to <t>E2</t> synthesis in MCF7 cells. Cells (1 × 10 6 per dish) were plated in 100-mm dishes 24h before treatments. Cells were then treated without or with panobinostat (10nM) for 0-180min (A-C) or other HDAC inhibitors for 45min (D-F). Following treatments, cells were collected, extracted with lysis buffer, and processed for either immunoprecipitation or immunoblotting studies, as described in Materials and methods . Representative immunoblots illustrate acetylation (Ac-StAR) and expression of StAR (T-StAR) in different treatment groups. IgG heavy chain (IgG-Hc) and β-actin expression were assessed for loading controls in immunoprecipitation and immunoblotting, respectively. Integrated optical density (IOD) values of Ac-StAR and T-StAR in each band were quantified and normalized with corresponding IgG-Hc and β-actin expression, and presented as fold response (C,F). E2 levels in media at each time point were determined by <t>ELISA</t> and presented as pg/mg protein (C). Data are representative of 3-4 independent experiments. *, p
    Elisa Kit, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 94/100, based on 1503 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kit/product/Cayman Chemical
    Average 94 stars, based on 1503 article reviews
    Price from $9.99 to $1999.99
    elisa kit - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Abcam phosphate buffered saline pbs
    Transforming growth factor-β (TGF-β) induces the colorectal cancer cells metastasis through the epithelial to mesenchymal transition progress. (A) Imagesof CT26 and HCT116 cells treated with phosphate buffered saline <t>(PBS),</t> TGF-β (5 ng/mL), tumor-associated macrophages (TAMs) cultural supernatant and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours. Scale bar=30 μm. (B) Relative <t>E-cadherin</t> expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (C) Relative vimentin expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (D) Western blotting of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for24 hours (E) Immunofluorescence of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (F) Immunofluorescence of E-cadherin andvimentin in tumor tissues of non-metastatic colorectal cancer patients (patients/N) andmetastatic colorectal cancer patients (patients/M). Scalebar=20 μm. The data was presented as the mean±standard error of mean from three independent experiments. *p
    Phosphate Buffered Saline Pbs, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2966 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphate buffered saline pbs/product/Abcam
    Average 99 stars, based on 2966 article reviews
    Price from $9.99 to $1999.99
    phosphate buffered saline pbs - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Abcam elisa kits
    Antitumor potency of the SA-IL-2 MCSCs vaccine in the pulmonary model mice. a In the cytotoxicity assay, the portion of CTL in the experimental group was significantly higher than in the four control groups. b In <t>ELISA</t> assay, the expression of serum <t>IgG</t> antibodies in the experimental group was significantly higher than in the four control groups. c FCM analysis showed that the portion of DCs (CD11c + CD80 + ) in the experimental group was significantly larger than in the four control groups. d FCM analysis showed that the portion of CD8 + and CD4 + T cells in the experimental group was significantly larger than in the four control groups. *P
    Elisa Kits, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1066 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kits/product/Abcam
    Average 99 stars, based on 1066 article reviews
    Price from $9.99 to $1999.99
    elisa kits - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Crystal Chem Inc ultra sensitive mouse insulin elisa kit
    Impaired insulin secretion from isolated Pick1 cKO islets. (a) <t>GSIS</t> was performed on isolated islets stimulated with 4.8 or 30 mM KCl or 2.8 or 16.7 mM glucose (Glc). (b) The stimulated index was also calculated following KCl or Glc treatment. (c) Insulin content in the isolated islets was measured by <t>ELISA.</t> (d) Isolated WT and Pick1 cKO islets were incubated in the presence of 16.7 mM Glc at 2-min intervals for 20 min. (e) Same data as shown in d, but plotted as release per 2 min. (f, g) Secreted insulin was tested from isolated WT and Pick1 cKO islets incubated with or without E4. Data are represented as mean ± SEM, n = 3, three to four groups per type of mouse, 10–0 islets per group. * p
    Ultra Sensitive Mouse Insulin Elisa Kit, supplied by Crystal Chem Inc, used in various techniques. Bioz Stars score: 93/100, based on 1190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ultra sensitive mouse insulin elisa kit/product/Crystal Chem Inc
    Average 93 stars, based on 1190 article reviews
    Price from $9.99 to $1999.99
    ultra sensitive mouse insulin elisa kit - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    94
    Millipore elisa kits
    Characterization of IFN-γ ( a , e ), IL-6 ( b , f ), <t>TNF-α</t> ( c , g ) and Lptn ( d , f ) secretion from the spleen ( a , b , c and d ) and lung ( a , b , c and d ) tissues of mice after CIV alone, S. pseudintermedius alone, or both. The concentrations in the supernatants of homogenates from the spleens and lungs on days 1, 3 and 5 post-challenge were measured by <t>ELISA.</t> The results are expressed in terms of pg/ml. Analysis of variance (ANOVA) was performed for the statistical analysis. *, P
    Elisa Kits, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1417 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kits/product/Millipore
    Average 94 stars, based on 1417 article reviews
    Price from $9.99 to $1999.99
    elisa kits - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    Recognition of wt rAAV2, rAAV8, and their chimeras by ADK8. (A) ADK8 ELISA for rAAV8 and rAAV8 with AAV2 amino acid substitution mutants. Wild-type rAAV8 and the mutants rAAV8 456GTANTQ461→GTTTQS and rAAV8 493TTTGQ497→KTSAD were detected by the ADK8, but the rAAV8 586LQQQNT591→LQRGNR mutant was not. (B) ADK8 ELISA for rAAV2, rAAV8, and rAAV2 with AAV8 amino acid substitution mutants. Wild-type rAAV2 and the rAAV2 457QSRLQ461→TQTLG mutant were not recognized by ADK8, but the rAAV2 585RGNRQ589→QQNTA mutant was. (C) ADK8 ELISA for rAAV8 and seven-residue peptide insertion (at aa 590) mutants. All insertions disrupted AAV8 detection by ADK8. In panels A to C, the mean standard deviations from four independent experiments are shown; asterisks identify mutants for which capsids could not be detected in the ADK8 ELISA (detection limit, 10 8 capsids/ml). A total of 10 10 viral genomes were used for each ELISA.

    Journal: Journal of Virology

    Article Title: Mapping a Neutralizing Epitope onto the Capsid of Adeno-Associated Virus Serotype 8

    doi: 10.1128/JVI.00218-12

    Figure Lengend Snippet: Recognition of wt rAAV2, rAAV8, and their chimeras by ADK8. (A) ADK8 ELISA for rAAV8 and rAAV8 with AAV2 amino acid substitution mutants. Wild-type rAAV8 and the mutants rAAV8 456GTANTQ461→GTTTQS and rAAV8 493TTTGQ497→KTSAD were detected by the ADK8, but the rAAV8 586LQQQNT591→LQRGNR mutant was not. (B) ADK8 ELISA for rAAV2, rAAV8, and rAAV2 with AAV8 amino acid substitution mutants. Wild-type rAAV2 and the rAAV2 457QSRLQ461→TQTLG mutant were not recognized by ADK8, but the rAAV2 585RGNRQ589→QQNTA mutant was. (C) ADK8 ELISA for rAAV8 and seven-residue peptide insertion (at aa 590) mutants. All insertions disrupted AAV8 detection by ADK8. In panels A to C, the mean standard deviations from four independent experiments are shown; asterisks identify mutants for which capsids could not be detected in the ADK8 ELISA (detection limit, 10 8 capsids/ml). A total of 10 10 viral genomes were used for each ELISA.

    Article Snippet: Horseradish peroxidase (HRP)-labeled (ApD Serotech, Oxford, United Kingdom) ADK8 (1 μg/ml) antibodies were then applied and analyzed by an ELISA plate reader (Ascent FL; Thermo Labsystems, Egelsbach, Germany).

    Techniques: Enzyme-linked Immunosorbent Assay, Mutagenesis

    341G2 hIgG1 (h1) and hIgG4 (h4) Suppress Immune Function In Vitro and In Vivo (A) Ramos cells were incubated with fixed concentration of CD40L and various concentrations of anti-CD40 mAbs. Remaining bound CD40L was detected by anti-FLAG-APC. Means ± SEM, n = 3, data representative of three experiments. (B) Purified hCD40Tg mouse splenic B cells were incubated with 2 μg/mL CD40L in the presence or absence of 5 μg/mL 341G2 h1 and h4 for 2 days. Cell culture images were taken on day 2. Proliferation was measured by 3 H-thymidine incorporation. Means ± SEM, n = 5, data representative of three experiments. Scale bar, 0.5 mm. (C) Purified human B cells were incubated with 2 μg/mL CD40L in the presence or absence of 5 μg/mL 341G2 h1 or h4 for 2 days. Cell culture images were taken on day 2. Proliferation was measured by 3 H-thymidine incorporation. Means ± SEM, n = 3–5, data representative of three donors. Scale bar, 0.5 mm. (D) hCD40Tg mice received 500 μg OVA and 100 μg anti-CD40 mAbs on day 0 and another dose of 100 μg anti-CD40 mAbs on day 3. Mice were bled on day 18 and serum levels of anti-OVA IgG were quantified by ELISA as described in the STAR Methods . Means ± SEM, n = 4–5, data representative of two experiments. Each dot represents one mouse. (E) Mice received the same treatment as in (D). The level of circulating CD19 + B cells in blood on day 2 was quantified by anti-mouse CD19-APC and expressed as the percentage of CD45.2 + cells. Means ± SEM, n = 4–5, data representative of two experiments. Each dot represents one mouse. Two-tailed, non-paired Student’s t test, ∗ p

    Journal: Cancer Cell

    Article Title: Isotype Switching Converts Anti-CD40 Antagonism to Agonism to Elicit Potent Antitumor Activity

    doi: 10.1016/j.ccell.2020.04.013

    Figure Lengend Snippet: 341G2 hIgG1 (h1) and hIgG4 (h4) Suppress Immune Function In Vitro and In Vivo (A) Ramos cells were incubated with fixed concentration of CD40L and various concentrations of anti-CD40 mAbs. Remaining bound CD40L was detected by anti-FLAG-APC. Means ± SEM, n = 3, data representative of three experiments. (B) Purified hCD40Tg mouse splenic B cells were incubated with 2 μg/mL CD40L in the presence or absence of 5 μg/mL 341G2 h1 and h4 for 2 days. Cell culture images were taken on day 2. Proliferation was measured by 3 H-thymidine incorporation. Means ± SEM, n = 5, data representative of three experiments. Scale bar, 0.5 mm. (C) Purified human B cells were incubated with 2 μg/mL CD40L in the presence or absence of 5 μg/mL 341G2 h1 or h4 for 2 days. Cell culture images were taken on day 2. Proliferation was measured by 3 H-thymidine incorporation. Means ± SEM, n = 3–5, data representative of three donors. Scale bar, 0.5 mm. (D) hCD40Tg mice received 500 μg OVA and 100 μg anti-CD40 mAbs on day 0 and another dose of 100 μg anti-CD40 mAbs on day 3. Mice were bled on day 18 and serum levels of anti-OVA IgG were quantified by ELISA as described in the STAR Methods . Means ± SEM, n = 4–5, data representative of two experiments. Each dot represents one mouse. (E) Mice received the same treatment as in (D). The level of circulating CD19 + B cells in blood on day 2 was quantified by anti-mouse CD19-APC and expressed as the percentage of CD45.2 + cells. Means ± SEM, n = 4–5, data representative of two experiments. Each dot represents one mouse. Two-tailed, non-paired Student’s t test, ∗ p

    Article Snippet: To detect anti-OVA IgG in serum, ELISA plates (Thermo Fisher Scientific) were coated with 5 μg/mL OVA in PBS overnight.

    Techniques: In Vitro, In Vivo, Incubation, Concentration Assay, Purification, Cell Culture, Mouse Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

    Efferocytosis of apoptotic mesenchymal stromal cells polarizes monocytes into an immunosuppressive phenotype. (A) Apoptotic mesenchymal stromal cells (ApoMSC) were stained with 20 ng/mL pHrodo™ Red-succinimidyl ester (PE) while isolated monocytes were stained with 1 μM CellTrace™ Violet (V450). Monocytes were cultured with different amounts of ApoMSC for 2 h at 37°C, 5% CO2. Lipopolysaccharide (LPS) 100 ng/mL was used to stimulate monocytes. The mean fluorescence intensity (MFI) (PE) within the monocyte population was measured as an indicator of efferocytosis, n=3. (B) MFI (PE) after 2 and 24 h was compared among the different groups, n=3. (C) Confocal images of monocytes efferocytosing ApoMSC after 24 h in culture. The yellow arrows indicate the monocytes engulfing ApoMSC while the white arrows indicate ApoMSC alone. The bar represents 20 μm. (D) Monocytes (2 × 10 5 /well) were co-cultured with ApoMSC (4 × 10 5 /well) for 8 h. CellTrace™ Violet-labeled CD3 T cells (2 × 10 5 /well) were added to the cultures and stimulated by CD3/CD28 beads at a 1:1 cell:bead ratio for 3 days. The percentages of proliferating T cells were measured by flow cytometry according to the loss of fluorescence intensity due to cell division, n=4. (E-G) Intracellular and surface staining was performed to examine the expression of different proteins, including COX2, IDO and PD-L1, in monocytes. The COX2 high , n=3 (E), IDO high , n=4 (F) and PD-L1 high , n=3 (G) populations were gated according to the monocyte alone control (used as a negative control). (H-J) Enzyme-linked immunosorbent assays were performed to assess the amount of PGE2, n=4 (H), IL-10, n=3 (I) and TNF-α, n=6 (J) in cell culture supernatants. (K) To select the monocytes according to efferocytosis, ApoMSC were fluorescent-labeled with CellTrace™ FarRed (APC + ) while monocytes were labeled with CellTrace™ Violet (V450 + ). APC + monocytes were regarded as efferocytosis + and APC − monocytes were regarded as efferocytosis − . (L) Bar charts showing the difference of COX2 high , IDO high and PD-L1 high populations between the efferocytosis + and efferocytosis- monocytes, n=3. Experimental data are expressed as means ± standard deviation. An unpaired t test was used to compare the mean differences between two samples. One-way analysis of variance and the post-hoc Tukey test were used to compare the mean differences among the samples (* P

    Journal: Haematologica

    Article Title: Apoptotic mesenchymal stromal cells induce prostaglandin E2 in monocytes: implications for the monitoring of mesenchymal stromal cell activity

    doi: 10.3324/haematol.2018.214767

    Figure Lengend Snippet: Efferocytosis of apoptotic mesenchymal stromal cells polarizes monocytes into an immunosuppressive phenotype. (A) Apoptotic mesenchymal stromal cells (ApoMSC) were stained with 20 ng/mL pHrodo™ Red-succinimidyl ester (PE) while isolated monocytes were stained with 1 μM CellTrace™ Violet (V450). Monocytes were cultured with different amounts of ApoMSC for 2 h at 37°C, 5% CO2. Lipopolysaccharide (LPS) 100 ng/mL was used to stimulate monocytes. The mean fluorescence intensity (MFI) (PE) within the monocyte population was measured as an indicator of efferocytosis, n=3. (B) MFI (PE) after 2 and 24 h was compared among the different groups, n=3. (C) Confocal images of monocytes efferocytosing ApoMSC after 24 h in culture. The yellow arrows indicate the monocytes engulfing ApoMSC while the white arrows indicate ApoMSC alone. The bar represents 20 μm. (D) Monocytes (2 × 10 5 /well) were co-cultured with ApoMSC (4 × 10 5 /well) for 8 h. CellTrace™ Violet-labeled CD3 T cells (2 × 10 5 /well) were added to the cultures and stimulated by CD3/CD28 beads at a 1:1 cell:bead ratio for 3 days. The percentages of proliferating T cells were measured by flow cytometry according to the loss of fluorescence intensity due to cell division, n=4. (E-G) Intracellular and surface staining was performed to examine the expression of different proteins, including COX2, IDO and PD-L1, in monocytes. The COX2 high , n=3 (E), IDO high , n=4 (F) and PD-L1 high , n=3 (G) populations were gated according to the monocyte alone control (used as a negative control). (H-J) Enzyme-linked immunosorbent assays were performed to assess the amount of PGE2, n=4 (H), IL-10, n=3 (I) and TNF-α, n=6 (J) in cell culture supernatants. (K) To select the monocytes according to efferocytosis, ApoMSC were fluorescent-labeled with CellTrace™ FarRed (APC + ) while monocytes were labeled with CellTrace™ Violet (V450 + ). APC + monocytes were regarded as efferocytosis + and APC − monocytes were regarded as efferocytosis − . (L) Bar charts showing the difference of COX2 high , IDO high and PD-L1 high populations between the efferocytosis + and efferocytosis- monocytes, n=3. Experimental data are expressed as means ± standard deviation. An unpaired t test was used to compare the mean differences between two samples. One-way analysis of variance and the post-hoc Tukey test were used to compare the mean differences among the samples (* P

    Article Snippet: Enzyme-linked immunosorbent assays (all from Thermo Fisher Scientific) were used to examine the secreted soluble factors in culture supernatants.

    Techniques: Staining, Isolation, Cell Culture, Fluorescence, Labeling, Flow Cytometry, Cytometry, Expressing, Negative Control, Standard Deviation

    Oral administration of TEE decreased corticosterone and increased brain-derived neurotrophic factor (BDNF) in in aged hippocampus. The corticosterone ( A ) and BDNF ( B ) levels in the mouse hippocampus were measured using ELISA. Each bar represents the mean ± SEM for each group ( n = 9). The asterisks denote a significant difference (*, p

    Journal: Nutrients

    Article Title: Lycopersicon esculentum Extract Enhances Cognitive Function and Hippocampal Neurogenesis in Aged Mice

    doi: 10.3390/nu8110679

    Figure Lengend Snippet: Oral administration of TEE decreased corticosterone and increased brain-derived neurotrophic factor (BDNF) in in aged hippocampus. The corticosterone ( A ) and BDNF ( B ) levels in the mouse hippocampus were measured using ELISA. Each bar represents the mean ± SEM for each group ( n = 9). The asterisks denote a significant difference (*, p

    Article Snippet: The corticosterone and BDNF levels were quantified at 450 nm using an ELISA reader (Thermo Fisher Scientific Inc., Waltham, MA, USA) and were analyzed by a standard curve.

    Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay

    Interleukin (IL)-12 production by human peripheral blood mononuclear cells (PBMC) co-cultured with Lactobacillus casei ssp. casei . The cells were co-cultured with the indicated doses of live L. casei ssp. casei, L. paracasei ssp. paracasei or L. acidophilus for 48 h. IL-12 production was analysed by enzyme-linked immunosorbent assay (ELISA). Data are shown as the mean ± s.e.m. of three independent experiments.

    Journal: Clinical and Experimental Immunology

    Article Title: Natural killer cell activities of synbiotic Lactobacillus casei ssp. casei in conjunction with dextran

    doi: 10.1111/j.1365-2249.2005.02975.x

    Figure Lengend Snippet: Interleukin (IL)-12 production by human peripheral blood mononuclear cells (PBMC) co-cultured with Lactobacillus casei ssp. casei . The cells were co-cultured with the indicated doses of live L. casei ssp. casei, L. paracasei ssp. paracasei or L. acidophilus for 48 h. IL-12 production was analysed by enzyme-linked immunosorbent assay (ELISA). Data are shown as the mean ± s.e.m. of three independent experiments.

    Article Snippet: Culture supernatants were collected and analysed by enzyme-linked immunosorbent assay (ELISA) for secreted IL-12 ( e Bioscience, San Diego, CA, USA).

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    Antibody responses, virus titers, and inflammatory cytokine levels in BALF and lungs after challenge. Levels of IgG antibodies were determined in samples collected from mice at day 5 p.i. with A/Philippines/2/82 (H3N2) virus ( n = 5). (A) IgG antibody levels in BALF. The antibody level was determined by ELISA using M2e peptide as a coating antigen. Lung viral titers (B), IFN-γ (C), and IL-6 (D) cytokines in BALF were determined at day 5 p.i. Lung viral titers were determined by an egg infection assay. IFN-γ and IL-6 were determined by a cytokine ELISA. Data represent mean ± SEM. Statistical significances were determined by 1-way ANOVA. Asterisks indicate significant differences (* p

    Journal: PLoS ONE

    Article Title: Cross Protection against Influenza A Virus by Yeast-Expressed Heterologous Tandem Repeat M2 Extracellular Proteins

    doi: 10.1371/journal.pone.0137822

    Figure Lengend Snippet: Antibody responses, virus titers, and inflammatory cytokine levels in BALF and lungs after challenge. Levels of IgG antibodies were determined in samples collected from mice at day 5 p.i. with A/Philippines/2/82 (H3N2) virus ( n = 5). (A) IgG antibody levels in BALF. The antibody level was determined by ELISA using M2e peptide as a coating antigen. Lung viral titers (B), IFN-γ (C), and IL-6 (D) cytokines in BALF were determined at day 5 p.i. Lung viral titers were determined by an egg infection assay. IFN-γ and IL-6 were determined by a cytokine ELISA. Data represent mean ± SEM. Statistical significances were determined by 1-way ANOVA. Asterisks indicate significant differences (* p

    Article Snippet: Cytokines in BALF were assayed with ELISA kits in duplicate against a standard curve according to the manufacturers’ instructions (eBioscience, San Diego, CA).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Infection

    Effect of IFN-γ cell preconditioning (1 ng/ml for 24 h) singly and with HRV-14 or HRV-1b infection (TCID 50 10 2.5 for 90 mins) on sICAM-1 protein release (A) and sICAM-1 gene expression (B) in NHBE cells over the study period . sICAM-1 in cell culture supernatants was assayed using ELISA. Data are mean ± S.E. of three separate experiments (Fig 5A,*p

    Journal: Journal of Inflammation (London, England)

    Article Title: IFN-gamma regulation of ICAM-1 receptors in bronchial epithelial cells: soluble ICAM-1 release inhibits human rhinovirus infection

    doi: 10.1186/1476-9255-5-8

    Figure Lengend Snippet: Effect of IFN-γ cell preconditioning (1 ng/ml for 24 h) singly and with HRV-14 or HRV-1b infection (TCID 50 10 2.5 for 90 mins) on sICAM-1 protein release (A) and sICAM-1 gene expression (B) in NHBE cells over the study period . sICAM-1 in cell culture supernatants was assayed using ELISA. Data are mean ± S.E. of three separate experiments (Fig 5A,*p

    Article Snippet: sICAM-1 protein ELISA Soluble ICAM-1 assays were performed with a commercially available ELISA kit (BioSource International, California USA).

    Techniques: Infection, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

    Effect of protease inhibitors on sICAM-1 release (A) and mICAM-1 levels (B) . Cell cultures were pre-incubated with 1 complete™ mini tablet/10 ml of media; sICAM-1 in associated supernatants was assayed using ELISA. Data are mean ± S.E. of three separate experiments (Fig. 7A, *p

    Journal: Journal of Inflammation (London, England)

    Article Title: IFN-gamma regulation of ICAM-1 receptors in bronchial epithelial cells: soluble ICAM-1 release inhibits human rhinovirus infection

    doi: 10.1186/1476-9255-5-8

    Figure Lengend Snippet: Effect of protease inhibitors on sICAM-1 release (A) and mICAM-1 levels (B) . Cell cultures were pre-incubated with 1 complete™ mini tablet/10 ml of media; sICAM-1 in associated supernatants was assayed using ELISA. Data are mean ± S.E. of three separate experiments (Fig. 7A, *p

    Article Snippet: sICAM-1 protein ELISA Soluble ICAM-1 assays were performed with a commercially available ELISA kit (BioSource International, California USA).

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay

    Interleukin-6 concentration in wild-type MEFs and ATF3 KO cells. Wild-type MEFs and ATF3 KO cells were treated with 1, 2, 4, or 8 µM doxorubicin, for 24 h. The concentration of interleukin-6 (IL-6) in the cell culture supernatant was measured using an enzyme-linked immunosorbent assay kit. The IL-6 concentration was calculated using a standard curve. Experiments were independently performed 5 times. **P

    Journal: PLoS ONE

    Article Title: Doxorubicin Induces Cytotoxicity through Upregulation of pERK-Dependent ATF3

    doi: 10.1371/journal.pone.0044990

    Figure Lengend Snippet: Interleukin-6 concentration in wild-type MEFs and ATF3 KO cells. Wild-type MEFs and ATF3 KO cells were treated with 1, 2, 4, or 8 µM doxorubicin, for 24 h. The concentration of interleukin-6 (IL-6) in the cell culture supernatant was measured using an enzyme-linked immunosorbent assay kit. The IL-6 concentration was calculated using a standard curve. Experiments were independently performed 5 times. **P

    Article Snippet: Cytokine Analysis The concentration of each cytokine in the supernatant of the culture media and serum was determined using commercially available enzyme-linked immunosorbent assay (ELISA) kits (eBioscience).

    Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    Expression of macrophage-inflammatory protein-2 (MIP-2) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. MIP-2 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Journal: Clinical and Experimental Immunology

    Article Title: The volatile anaesthetic sevoflurane attenuates lipopolysaccharide-induced injury in alveolar macrophages

    doi: 10.1111/j.1365-2249.2008.03807.x

    Figure Lengend Snippet: Expression of macrophage-inflammatory protein-2 (MIP-2) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. MIP-2 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Article Snippet: Enzyme-linked immunosorbent assays were performed according to the manufacturer's protocol assessing the cytokine TNF-α (BD Biosciences, San Diego, CA, USA), the chemokines CINC-1 (R & D Systems Europe Ltd, Abingdon, UK), MIP-2 (R & D Systems Europe Ltd) and MCP-1 (BD Biosciences).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Expression of monocyte chemoattractant protein-1 (MCP-1) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. MCP-1 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Journal: Clinical and Experimental Immunology

    Article Title: The volatile anaesthetic sevoflurane attenuates lipopolysaccharide-induced injury in alveolar macrophages

    doi: 10.1111/j.1365-2249.2008.03807.x

    Figure Lengend Snippet: Expression of monocyte chemoattractant protein-1 (MCP-1) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. MCP-1 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Article Snippet: Enzyme-linked immunosorbent assays were performed according to the manufacturer's protocol assessing the cytokine TNF-α (BD Biosciences, San Diego, CA, USA), the chemokines CINC-1 (R & D Systems Europe Ltd, Abingdon, UK), MIP-2 (R & D Systems Europe Ltd) and MCP-1 (BD Biosciences).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Expression of cytokine-induced neutrophil chemoattractant-1 (CINC-1) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. CINC-1 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Journal: Clinical and Experimental Immunology

    Article Title: The volatile anaesthetic sevoflurane attenuates lipopolysaccharide-induced injury in alveolar macrophages

    doi: 10.1111/j.1365-2249.2008.03807.x

    Figure Lengend Snippet: Expression of cytokine-induced neutrophil chemoattractant-1 (CINC-1) under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by a co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. CINC-1 enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Article Snippet: Enzyme-linked immunosorbent assays were performed according to the manufacturer's protocol assessing the cytokine TNF-α (BD Biosciences, San Diego, CA, USA), the chemokines CINC-1 (R & D Systems Europe Ltd, Abingdon, UK), MIP-2 (R & D Systems Europe Ltd) and MCP-1 (BD Biosciences).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Expression of tumour necrosis factor (TNF)-α under stimulation with endotoxin and application of different concentrations of sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) for 2 h, followed by a co-application of sevoflurane (sevo) (1·1, 2·2, 3·3 and 4·4 vol%) or control gas for 4 h. TNF-α enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from four experiments. P

    Journal: Clinical and Experimental Immunology

    Article Title: The volatile anaesthetic sevoflurane attenuates lipopolysaccharide-induced injury in alveolar macrophages

    doi: 10.1111/j.1365-2249.2008.03807.x

    Figure Lengend Snippet: Expression of tumour necrosis factor (TNF)-α under stimulation with endotoxin and application of different concentrations of sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) for 2 h, followed by a co-application of sevoflurane (sevo) (1·1, 2·2, 3·3 and 4·4 vol%) or control gas for 4 h. TNF-α enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from four experiments. P

    Article Snippet: Enzyme-linked immunosorbent assays were performed according to the manufacturer's protocol assessing the cytokine TNF-α (BD Biosciences, San Diego, CA, USA), the chemokines CINC-1 (R & D Systems Europe Ltd, Abingdon, UK), MIP-2 (R & D Systems Europe Ltd) and MCP-1 (BD Biosciences).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Expression of tumour necrosis factor (TNF)-α under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. TNF-α enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Journal: Clinical and Experimental Immunology

    Article Title: The volatile anaesthetic sevoflurane attenuates lipopolysaccharide-induced injury in alveolar macrophages

    doi: 10.1111/j.1365-2249.2008.03807.x

    Figure Lengend Snippet: Expression of tumour necrosis factor (TNF)-α under stimulation with endotoxin and application of 3·3 vol% sevoflurane. Alveolar macrophages were stimulated with lipopolysaccharide (LPS) [control (co) with phosphate-buffered saline] for 2 h, followed by co-application of 3·3 vol% sevoflurane (sevo) (white bars) or control gas (black bars) for 2, 4, 6, 8, 12 and 24 h. TNF-α enzyme-linked immunosorbent assay was performed with supernatants. Values are mean ± standard error of the mean from three experiments. P

    Article Snippet: Enzyme-linked immunosorbent assays were performed according to the manufacturer's protocol assessing the cytokine TNF-α (BD Biosciences, San Diego, CA, USA), the chemokines CINC-1 (R & D Systems Europe Ltd, Abingdon, UK), MIP-2 (R & D Systems Europe Ltd) and MCP-1 (BD Biosciences).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Effects of EA on inflammatory cytokines levels in the peri-infarct hippocampal CA1and sensorimotor cortex of MCAO/R rats. a Representative and quantitative analysis of ELISA showed that EA treatment for 7 days decreased the levels of IL-1β and b increased the levels of IL-10 in MCAO/R injured rats. Data are presented as mean ± S.E.M. from 6 individual rats in each group. P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Electroacupuncture ameliorating post-stroke cognitive impairments via inhibition of peri-infarct astroglial and microglial/macrophage P2 purinoceptors-mediated neuroinflammation and hyperplasia

    doi: 10.1186/s12906-017-1974-y

    Figure Lengend Snippet: Effects of EA on inflammatory cytokines levels in the peri-infarct hippocampal CA1and sensorimotor cortex of MCAO/R rats. a Representative and quantitative analysis of ELISA showed that EA treatment for 7 days decreased the levels of IL-1β and b increased the levels of IL-10 in MCAO/R injured rats. Data are presented as mean ± S.E.M. from 6 individual rats in each group. P

    Article Snippet: The quantitative absorbance was determined at 450 nm using an ELISA reader (BioTek, Model ELX800, USA) according to IL-1β or IL-10 protein standards.

    Techniques: Enzyme-linked Immunosorbent Assay

    Antigenic analysis of Fwt and Flys-GCN. F protein preparations Fwt and Flys-GCN were coated in 96 well plates for ELISA analysis. The reactivity of the recombinant proteins with the different antibodies (indicated above each graph) was determined by applying 2-fold dilutions of these antibodies as indicated in the Materials and Methods. Binding of the antibodies was detected using HRP-conjugated secondary antibodies. The antibody dilution is indicated on the X-axes, while the Y-axes corresponds to optical density read at 450nm (OD450 nm). Error bars indicate standard deviations (n = 3).

    Journal: PLoS ONE

    Article Title: Recombinant Soluble Respiratory Syncytial Virus F Protein That Lacks Heptad Repeat B, Contains a GCN4 Trimerization Motif and Is Not Cleaved Displays Prefusion-Like Characteristics

    doi: 10.1371/journal.pone.0130829

    Figure Lengend Snippet: Antigenic analysis of Fwt and Flys-GCN. F protein preparations Fwt and Flys-GCN were coated in 96 well plates for ELISA analysis. The reactivity of the recombinant proteins with the different antibodies (indicated above each graph) was determined by applying 2-fold dilutions of these antibodies as indicated in the Materials and Methods. Binding of the antibodies was detected using HRP-conjugated secondary antibodies. The antibody dilution is indicated on the X-axes, while the Y-axes corresponds to optical density read at 450nm (OD450 nm). Error bars indicate standard deviations (n = 3).

    Article Snippet: Detection of HRP reactivity was performed using tetramethylbenzidine substrate (BioFX) and a ELISA plate reader (EL-808 from Biotek).

    Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay

    miR-146a, miR-335 and miR-622 regulate THP-1 cell migration, invasion and proliferation. (A–C) THP-1 cells were transiently transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Then, cells were loaded in Boyden chamber on uncoated filters (A and B) or Matrigel coated filters (C). Then, cells were allowed to migrate towards the uPA aminoterminal fragment (ATF) or the stroma derived factor 1 (SDF1) (A) or towards 10% serum (FBS) (B and C). Migrated cells were fixed, stained with haematoxylin, and counted. Results of migration assays are expressed as percentage of cells migrated towards chemoattractants over the cells migrated without chemoattractants; 100% values represent cell migration in the absence of chemoattractants. The values are the mean ± SEM of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test. (D) THP-1 cells were transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Cells, harvested at indicated times, were loaded in 96-well plates and incubated with 20 μl/well of CellTiter 96 AQueous One Solution Reagent for 4 hrs at 37°C, 5% CO 2 . Then, the absorbance was determined by an ELISA reader (Bio-Rad) at a wavelength of 490 nm. Fold increase represents the ratio between the OD at indicated time points and OD at time 0. The values are the mean of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Urokinase receptor and CXCR4 are regulated by common microRNAs in leukaemia cells

    doi: 10.1111/jcmm.12617

    Figure Lengend Snippet: miR-146a, miR-335 and miR-622 regulate THP-1 cell migration, invasion and proliferation. (A–C) THP-1 cells were transiently transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Then, cells were loaded in Boyden chamber on uncoated filters (A and B) or Matrigel coated filters (C). Then, cells were allowed to migrate towards the uPA aminoterminal fragment (ATF) or the stroma derived factor 1 (SDF1) (A) or towards 10% serum (FBS) (B and C). Migrated cells were fixed, stained with haematoxylin, and counted. Results of migration assays are expressed as percentage of cells migrated towards chemoattractants over the cells migrated without chemoattractants; 100% values represent cell migration in the absence of chemoattractants. The values are the mean ± SEM of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test. (D) THP-1 cells were transfected with the synthetic precursors of miR-146a, miR-335, miR-622 or of a control miR. Cells, harvested at indicated times, were loaded in 96-well plates and incubated with 20 μl/well of CellTiter 96 AQueous One Solution Reagent for 4 hrs at 37°C, 5% CO 2 . Then, the absorbance was determined by an ELISA reader (Bio-Rad) at a wavelength of 490 nm. Fold increase represents the ratio between the OD at indicated time points and OD at time 0. The values are the mean of three experiments performed in triplicate. (*) P ≤ 0.05, as determined by the Student’s t -test.

    Article Snippet: After incubation for 4 hrs at 37°C, 5% CO2 , the absorbance was determined by an ELISA reader (Bio-Rad) at a wavelength of 490 nm.

    Techniques: Migration, Transfection, Derivative Assay, Staining, Incubation, Enzyme-linked Immunosorbent Assay

    Effects of ASWP and three ASEPs on cell viability; phagocytic activity; and the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-12 in RAW264.7 macrophages. ASWP was obtained from Aster scaber using hot water extraction and ASEP-A, ASEP-C, and ASEP-P were obtained from Aster scaber using α-amylase, cellulase, and pectinase, respectively, in an enzyme-assisted extraction. RAW264.7 macrophages were stimulated with the polysaccharide samples at 1, 10, or 100 μg/mL. LPS (1 μg/mL) was used as the positive control (PC). ( A ) Cell viability was determined by the cell counting kit-8 (CCK-8) assay; ( B ) Phagocytic activity was determined using a phagocytosis assay; ( C ) NO levels in the culture media were determined by measuring nitrite accumulation, and the secretion levels of TNF-α, IL-6, and IL-12 were measured by ELISA. ( D ) mRNA expression levels of iNOS, TNF-α, IL-6, and IL-12 in macrophages were quantified by qRT-PCR analysis. Lowercase letters (a–c) indicate significant differences ( p

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of Pectinase-Assisted Extraction on the Physicochemical and Biological Properties of Polysaccharides from Aster scaber

    doi: 10.3390/ijms19092839

    Figure Lengend Snippet: Effects of ASWP and three ASEPs on cell viability; phagocytic activity; and the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-12 in RAW264.7 macrophages. ASWP was obtained from Aster scaber using hot water extraction and ASEP-A, ASEP-C, and ASEP-P were obtained from Aster scaber using α-amylase, cellulase, and pectinase, respectively, in an enzyme-assisted extraction. RAW264.7 macrophages were stimulated with the polysaccharide samples at 1, 10, or 100 μg/mL. LPS (1 μg/mL) was used as the positive control (PC). ( A ) Cell viability was determined by the cell counting kit-8 (CCK-8) assay; ( B ) Phagocytic activity was determined using a phagocytosis assay; ( C ) NO levels in the culture media were determined by measuring nitrite accumulation, and the secretion levels of TNF-α, IL-6, and IL-12 were measured by ELISA. ( D ) mRNA expression levels of iNOS, TNF-α, IL-6, and IL-12 in macrophages were quantified by qRT-PCR analysis. Lowercase letters (a–c) indicate significant differences ( p

    Article Snippet: Meanwhile, the secretion of TNF-α, IL-6, and IL-12 from the macrophages were assessed using corresponding ELISA kits (BD Biosciences, Pharmingen, San Diego, CA, USA).

    Techniques: Activity Assay, Positive Control, Cell Counting, CCK-8 Assay, Phagocytosis Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

    UPS13 is essential for Paxillin-mediated NLRP3 deubiquitination upon ATP treatment. A HEK293T cells were co-transfected with pHA-Paxillin and pFlag-BRCC3, pFlag-EIF3S5, pFlag-UPS13 or pFlag-OTUB1. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). B Hela cells stably expressing sh-UPS13 or sh-EIF3S5 were generated and analyzed. C The stable Hela cells were co-transfected with pFlag-NLRP3, pHA-UB, or pMyc-Paxillin. Lysates were prepared and subjected to denature-IP (top) or subjected to Western blot (bottom). D, E HEK293T cells were transfected with pFlag-UPS13 and pHA-Paxillin. F, G HEK293T cells were transfected with pFlag-UPS13 and pHA-NLRP3. Lysates were prepared and subjected to IP (top) or subjected to Western blot (bottom) (D–G). H HEK293T cells were co-transfected with pFlag-UPS13 and pHA-NLRP3 or pHA-NLRP3 mutants. I The diagrams of UPS13 and its mutants, as indicated. J HEK293T cells were co-transfected with pHA-NLRP3 and pFlag-UPS13 or pFlag-UPS13 mutants. K HEK293T cells were co-transfected with pHA-Paxillin and pFlag-UPS13 or pFlag-UPS13 mutants. L HEK293T cells were co-transfected with pFlag-UPS13 and pHA-Paxillin or pHA-Paxillin mutants. Lysates were prepared and subjected to IP (top) or subjected to Western blot (bottom) (H, j –l). M HEK293T cells stably expressing sh-UPS13 were generated. The stable cells were co-transfected with pFlag-NLRP3 and pHA-UB. N The stable HEK293T cells were co-transfected with pFlag-NLRP3, or pHA-UB with or without Spautin-1 (20 μM) for 6 h. O The stable HEK293T cells wre co-transfected with pFlag-NLRP3, pHA-UB, or pMyc-Paxillin with or without Spautin-1 (20 μM) for 6 h. Lysates were prepared and subjected to denature-IP (top) or subjected to Western blot (bottom) (m–o). P , Q TPA-differentiated THP-1 macrophages were treated with ATP (5 mM) for 2 h with/without Spautin-1 (5, 10, and 20 μM). IL-1β in supernatants was determined by ELISA (P). Mature IL-1β(p17) and cleaved Caspase-1(p22/p20) in supernatants and Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (Q). Data shown are means ± SEMs; ***p

    Journal: bioRxiv

    Article Title: Paxillin Promotes ATP-induced Activation of P2X7 Receptor and NLRP3 Inflammasome

    doi: 10.1101/2020.04.03.023721

    Figure Lengend Snippet: UPS13 is essential for Paxillin-mediated NLRP3 deubiquitination upon ATP treatment. A HEK293T cells were co-transfected with pHA-Paxillin and pFlag-BRCC3, pFlag-EIF3S5, pFlag-UPS13 or pFlag-OTUB1. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). B Hela cells stably expressing sh-UPS13 or sh-EIF3S5 were generated and analyzed. C The stable Hela cells were co-transfected with pFlag-NLRP3, pHA-UB, or pMyc-Paxillin. Lysates were prepared and subjected to denature-IP (top) or subjected to Western blot (bottom). D, E HEK293T cells were transfected with pFlag-UPS13 and pHA-Paxillin. F, G HEK293T cells were transfected with pFlag-UPS13 and pHA-NLRP3. Lysates were prepared and subjected to IP (top) or subjected to Western blot (bottom) (D–G). H HEK293T cells were co-transfected with pFlag-UPS13 and pHA-NLRP3 or pHA-NLRP3 mutants. I The diagrams of UPS13 and its mutants, as indicated. J HEK293T cells were co-transfected with pHA-NLRP3 and pFlag-UPS13 or pFlag-UPS13 mutants. K HEK293T cells were co-transfected with pHA-Paxillin and pFlag-UPS13 or pFlag-UPS13 mutants. L HEK293T cells were co-transfected with pFlag-UPS13 and pHA-Paxillin or pHA-Paxillin mutants. Lysates were prepared and subjected to IP (top) or subjected to Western blot (bottom) (H, j –l). M HEK293T cells stably expressing sh-UPS13 were generated. The stable cells were co-transfected with pFlag-NLRP3 and pHA-UB. N The stable HEK293T cells were co-transfected with pFlag-NLRP3, or pHA-UB with or without Spautin-1 (20 μM) for 6 h. O The stable HEK293T cells wre co-transfected with pFlag-NLRP3, pHA-UB, or pMyc-Paxillin with or without Spautin-1 (20 μM) for 6 h. Lysates were prepared and subjected to denature-IP (top) or subjected to Western blot (bottom) (m–o). P , Q TPA-differentiated THP-1 macrophages were treated with ATP (5 mM) for 2 h with/without Spautin-1 (5, 10, and 20 μM). IL-1β in supernatants was determined by ELISA (P). Mature IL-1β(p17) and cleaved Caspase-1(p22/p20) in supernatants and Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (Q). Data shown are means ± SEMs; ***p

    Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of human IL-1β in culture supernatants were measured by ELISA kit (BD Biosciences, San Jose, CA, USA).

    Techniques: Transfection, Western Blot, Stable Transfection, Expressing, Generated, Enzyme-linked Immunosorbent Assay

    ATP promotes Paxillin phosphorylation and Paxillin-NLRP3 interaction. A–D THP-1 cells stably expressing control lentivirus or Paxillin lentivirus were generated and differentiated to macrophages, which were then treated with ATP (5 mM) or Nigericin (2 μM) for 2 h (A, B), treated with DMSO, YVAD (50 μM), Glybenclamide (25 μg/ml), A438079 (100 μM), AZ10606120 (100 μM), and BAPTA-AM (30 μM) for 1 h before the treatment with ATP (5 mM) for 2 h (C, D). IL-1β levels in supernatants were determined by ELISA (A, C). Mature IL-1β (p17) and cleaved Casp-1 (p22/p20) in supernatants or Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (B, D). E TPA-differentiated THP-1 macrophages were treated with DMSO or ATP (5 mM) for 2 h. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). F BMDM cells prepared from C57BL/6 mice bone marrow were treated with LPS (1 μg/ml) for 6 h, and the primed BMDMs were stimulated by DMSO or ATP (5 mM) for 30 min. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). G, H TPA-differentiated THP-1 macrophages were treated with ATP (5 mM) for 30, 60, and 120 min (G). LPS-primed BMDMs were stimulated by ATP (5 mM) for 5, 15, and 30 min (H). The protein level of p-Paxillin(Y 118), p-Paxillin(Y31), Paxillin, and GAPDH was determined by Western blot. I HEK293T cells were co-transfected with pFlag-NLRP3 and pHA-Vector, pHA-Paxillin, pHA-Paxillin-(Y31A), and pHA-Paxillin-(Y118A). Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). J TPA-differentiated THP-1 macrophages were treated with DMSO, A438079 (100 μM), AZ10606120 (100 μM) for 1 h before the treatment with ATP (5 mM) for 2 h. The protein level of p-Paxillin(Y118), Paxillin, and GAPDH was determined by Western blot. Data shown are means ± SEMs; ***p

    Journal: bioRxiv

    Article Title: Paxillin Promotes ATP-induced Activation of P2X7 Receptor and NLRP3 Inflammasome

    doi: 10.1101/2020.04.03.023721

    Figure Lengend Snippet: ATP promotes Paxillin phosphorylation and Paxillin-NLRP3 interaction. A–D THP-1 cells stably expressing control lentivirus or Paxillin lentivirus were generated and differentiated to macrophages, which were then treated with ATP (5 mM) or Nigericin (2 μM) for 2 h (A, B), treated with DMSO, YVAD (50 μM), Glybenclamide (25 μg/ml), A438079 (100 μM), AZ10606120 (100 μM), and BAPTA-AM (30 μM) for 1 h before the treatment with ATP (5 mM) for 2 h (C, D). IL-1β levels in supernatants were determined by ELISA (A, C). Mature IL-1β (p17) and cleaved Casp-1 (p22/p20) in supernatants or Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (B, D). E TPA-differentiated THP-1 macrophages were treated with DMSO or ATP (5 mM) for 2 h. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). F BMDM cells prepared from C57BL/6 mice bone marrow were treated with LPS (1 μg/ml) for 6 h, and the primed BMDMs were stimulated by DMSO or ATP (5 mM) for 30 min. Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). G, H TPA-differentiated THP-1 macrophages were treated with ATP (5 mM) for 30, 60, and 120 min (G). LPS-primed BMDMs were stimulated by ATP (5 mM) for 5, 15, and 30 min (H). The protein level of p-Paxillin(Y 118), p-Paxillin(Y31), Paxillin, and GAPDH was determined by Western blot. I HEK293T cells were co-transfected with pFlag-NLRP3 and pHA-Vector, pHA-Paxillin, pHA-Paxillin-(Y31A), and pHA-Paxillin-(Y118A). Lysates were prepared and subjected to IP (top) or subjected to Western blot (as input) (bottom). J TPA-differentiated THP-1 macrophages were treated with DMSO, A438079 (100 μM), AZ10606120 (100 μM) for 1 h before the treatment with ATP (5 mM) for 2 h. The protein level of p-Paxillin(Y118), Paxillin, and GAPDH was determined by Western blot. Data shown are means ± SEMs; ***p

    Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of human IL-1β in culture supernatants were measured by ELISA kit (BD Biosciences, San Jose, CA, USA).

    Techniques: Stable Transfection, Expressing, Generated, Enzyme-linked Immunosorbent Assay, Western Blot, Mouse Assay, Transfection, Plasmid Preparation

    Paxillin is required for ATP- and Nigericin-induced NLRP3 inflammasome activation. A, B BMDMs prepared from C57BL/6 mice bone marrow were infected by lentivirus that expressing sh-Paxillin for 3 days. LPS-primed BMDMs were stimulated by MDP (50 μg/ml) for 6 h, ATP (5 mM) for 30 min, poly(dA:dT) (5 μg/ml) for 6 h, or Salmonella for 4 h. C, D LPS-primed BMDMs were stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, MSU (100 μg/ml) for 6 h, or Alum crystals (200 mg/ml) for 6 h. E, F BMDCs prepared from C57BL/6 mice bone marrow were infected by lentivirus that express sh-Paxillin for 3 days. LPS-primed BMDCs were stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, or MSU (100 μg/ml) for 6 h. IL-1β levels in supernatants were determined by ELISA (A, C, and E). Mature IL-1β (p17) and cleaved Casp-1 (p10) in supernatants as well as Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (B, D, and F). G, H PBMCs isolated from healthy individuals were infected by lentivirus that expresses sh-Paxillin for 2 days. Before stimulation, PBMCs were treated with LPS (1 μg/ml) for 6 h, and PBMCs were then stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, MSU (100 μg/ml) for 6 h, Alum crystals (200 mg/ml) for 6 h, or MDP (50 μg/ml) for 6 h. Paxillin in lysates was determined by Western blot (G). IL-1β levels in supernatants were determined by ELISA (H). I, J THP-1 cells stably expressing sh-NC or sh-Paxillin were generated and differentiated to macrophages, which were then treated with ATP (5 mM) for 2 h, Nigericin (2 μM) for 2 h, or MSU (100 μg/ml) for 6 h. IL-1β levels in supernatants were determined by ELISA (I). Mature IL-1β (p17) and cleaved Casp-1 (p22/p20) in supernatants and Paxillin, pro-IL-1β and pro-Casp-1 in lysates were determined by Western blot (J). Data shown are means ± SEMs; ***p

    Journal: bioRxiv

    Article Title: Paxillin Promotes ATP-induced Activation of P2X7 Receptor and NLRP3 Inflammasome

    doi: 10.1101/2020.04.03.023721

    Figure Lengend Snippet: Paxillin is required for ATP- and Nigericin-induced NLRP3 inflammasome activation. A, B BMDMs prepared from C57BL/6 mice bone marrow were infected by lentivirus that expressing sh-Paxillin for 3 days. LPS-primed BMDMs were stimulated by MDP (50 μg/ml) for 6 h, ATP (5 mM) for 30 min, poly(dA:dT) (5 μg/ml) for 6 h, or Salmonella for 4 h. C, D LPS-primed BMDMs were stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, MSU (100 μg/ml) for 6 h, or Alum crystals (200 mg/ml) for 6 h. E, F BMDCs prepared from C57BL/6 mice bone marrow were infected by lentivirus that express sh-Paxillin for 3 days. LPS-primed BMDCs were stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, or MSU (100 μg/ml) for 6 h. IL-1β levels in supernatants were determined by ELISA (A, C, and E). Mature IL-1β (p17) and cleaved Casp-1 (p10) in supernatants as well as Paxillin, pro-IL-1β, and pro-Casp-1 in lysates were determined by Western blot (B, D, and F). G, H PBMCs isolated from healthy individuals were infected by lentivirus that expresses sh-Paxillin for 2 days. Before stimulation, PBMCs were treated with LPS (1 μg/ml) for 6 h, and PBMCs were then stimulated by ATP (5 mM) for 30 min, Nigericin (2 μM) for 2 h, MSU (100 μg/ml) for 6 h, Alum crystals (200 mg/ml) for 6 h, or MDP (50 μg/ml) for 6 h. Paxillin in lysates was determined by Western blot (G). IL-1β levels in supernatants were determined by ELISA (H). I, J THP-1 cells stably expressing sh-NC or sh-Paxillin were generated and differentiated to macrophages, which were then treated with ATP (5 mM) for 2 h, Nigericin (2 μM) for 2 h, or MSU (100 μg/ml) for 6 h. IL-1β levels in supernatants were determined by ELISA (I). Mature IL-1β (p17) and cleaved Casp-1 (p22/p20) in supernatants and Paxillin, pro-IL-1β and pro-Casp-1 in lysates were determined by Western blot (J). Data shown are means ± SEMs; ***p

    Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of human IL-1β in culture supernatants were measured by ELISA kit (BD Biosciences, San Jose, CA, USA).

    Techniques: Activation Assay, Mouse Assay, Infection, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Isolation, Stable Transfection, Generated

    Effect of curcumin on A549 cell viability. The cytotoxic effects of curcumin on A549 cells were examined by MTT assay. Cells were cultured in growth medium at different concentrations (0–80 μM) for 12 h and 24 h, and absorbance at 540 nm was measured using an ELISA reader. Bar graphs indicate the percentage of viability. All data are expressed as mean ± SEM of three independent experiments. *** p

    Journal: International Journal of Molecular Sciences

    Article Title: Transcriptional Activation of Human GD3 Synthase (hST8Sia I) Gene in Curcumin-Induced Autophagy in A549 Human Lung Carcinoma Cells

    doi: 10.3390/ijms19071943

    Figure Lengend Snippet: Effect of curcumin on A549 cell viability. The cytotoxic effects of curcumin on A549 cells were examined by MTT assay. Cells were cultured in growth medium at different concentrations (0–80 μM) for 12 h and 24 h, and absorbance at 540 nm was measured using an ELISA reader. Bar graphs indicate the percentage of viability. All data are expressed as mean ± SEM of three independent experiments. *** p

    Article Snippet: The cells were then reacted with the MTT assay mixture for 30 min. DMSO treatment was then performed to dissolve the reduced formazan crystal from MTT, and the absorbance at 540 nm was measured using the ELISA plate reader (Bio-rad, Hercules, CA, USA).

    Techniques: MTT Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    Detection of secreted albumin and blood urea nitrogen by ELISA. The ALB and BUN concentration of UC-MSCs and BM-MSCs culture supernatants collected on weeks 1, 2, 3, 4 and 5 following hepatic differentiation were measured. The medium of hepatocyte was measured as a control. Each bar represents the mean ± standard deviation (n=3). *P

    Journal: Molecular Medicine Reports

    Article Title: Differentiation of umbilical cord mesenchymal stem cells into hepatocytes in comparison with bone marrow mesenchymal stem cells

    doi: 10.3892/mmr.2018.9181

    Figure Lengend Snippet: Detection of secreted albumin and blood urea nitrogen by ELISA. The ALB and BUN concentration of UC-MSCs and BM-MSCs culture supernatants collected on weeks 1, 2, 3, 4 and 5 following hepatic differentiation were measured. The medium of hepatocyte was measured as a control. Each bar represents the mean ± standard deviation (n=3). *P

    Article Snippet: ALB and blood urea nitrogen (BUN) contents were measured using ELISA kit (Human Albumin ELISA kit ab108788 and Bmassay, Human Blood Ureas Nitrogen ELISA kit 27013; Abcam, Cambridge, UK) according to the manufacturer's instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation

    The synthesized GILZ-p decreased LPS induced inflammatory cytokine secretion in culture medium of Müller cells. The Enzyme-Linked Immunosorbent Assays (ELISA) were performed to determine the protein expression levels of IL-1β, MCP-1, TNF-α, and ICAM-1 in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 6 for each group. ∗ P

    Journal: Frontiers in Pharmacology

    Article Title: A Synthesized Glucocorticoid- Induced Leucine Zipper Peptide Inhibits Retinal Müller Cell Gliosis

    doi: 10.3389/fphar.2018.00331

    Figure Lengend Snippet: The synthesized GILZ-p decreased LPS induced inflammatory cytokine secretion in culture medium of Müller cells. The Enzyme-Linked Immunosorbent Assays (ELISA) were performed to determine the protein expression levels of IL-1β, MCP-1, TNF-α, and ICAM-1 in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 6 for each group. ∗ P

    Article Snippet: Enzyme-Linked Immunosorbent Assays The culture supernatant concentrations of TNF-α, ICAM-1, MCP-1, and IL-1βwere measured after stimulation of Müller cells using ELISA kits (TNF alpha Rat ELISA Kit, ab46070, Abcam; Rat ICAM-1 ELISA Kit, EK0372, BOSTER; MCP-1 Rat ELISA Kit, ab100778, Abcam; and Interleukin-1 Beta Rat ELISA Kit, ab100768, Abcam; respectively).

    Techniques: Synthesized, Enzyme-linked Immunosorbent Assay, Expressing, MANN-WHITNEY

    Effect of Imm124-E on serum levels of IL-6, GLP-1, and adiponectin. All sera were measured using ELISA kits at days 1 and 30 of the trial in all treated patients. Graphs show ( A ) Serum levels of IL-6 from six patients; ( B ) Serum levels of GLP-1 post–glucose tolerance test from five patients; ( C ) Serum levels of adiponectin from eight patients. Note: Graphs indicate means ± SD. Abbreviations: IL-6, interleukin-6; GLP-1, glucagon-like peptide 1; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation.

    Journal: Journal of Inflammation Research

    Article Title: Alleviation of insulin resistance and liver damage by oral administration of Imm124-E is mediated by increased Tregs and associated with increased serum GLP-1 and adiponectin: results of a phase I/II clinical trial in NASH

    doi: 10.2147/JIR.S35227

    Figure Lengend Snippet: Effect of Imm124-E on serum levels of IL-6, GLP-1, and adiponectin. All sera were measured using ELISA kits at days 1 and 30 of the trial in all treated patients. Graphs show ( A ) Serum levels of IL-6 from six patients; ( B ) Serum levels of GLP-1 post–glucose tolerance test from five patients; ( C ) Serum levels of adiponectin from eight patients. Note: Graphs indicate means ± SD. Abbreviations: IL-6, interleukin-6; GLP-1, glucagon-like peptide 1; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation.

    Article Snippet: The circulating level of human GLP-1 was quantified using a commercial ELISA kit from Millipore Corp (Billerica, MA) according to the manufacturer’s instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

    Acetylation and expression of StAR in response to HDAC inhibitors, and their correlation to E2 synthesis in MCF7 cells. Cells (1 × 10 6 per dish) were plated in 100-mm dishes 24h before treatments. Cells were then treated without or with panobinostat (10nM) for 0-180min (A-C) or other HDAC inhibitors for 45min (D-F). Following treatments, cells were collected, extracted with lysis buffer, and processed for either immunoprecipitation or immunoblotting studies, as described in Materials and methods . Representative immunoblots illustrate acetylation (Ac-StAR) and expression of StAR (T-StAR) in different treatment groups. IgG heavy chain (IgG-Hc) and β-actin expression were assessed for loading controls in immunoprecipitation and immunoblotting, respectively. Integrated optical density (IOD) values of Ac-StAR and T-StAR in each band were quantified and normalized with corresponding IgG-Hc and β-actin expression, and presented as fold response (C,F). E2 levels in media at each time point were determined by ELISA and presented as pg/mg protein (C). Data are representative of 3-4 independent experiments. *, p

    Journal: Biochemical and biophysical research communications

    Article Title: Overexpression of the steroidogenic acute regulatory protein in breast cancer: Regulation by histone deacetylase inhibition

    doi: 10.1016/j.bbrc.2018.12.145

    Figure Lengend Snippet: Acetylation and expression of StAR in response to HDAC inhibitors, and their correlation to E2 synthesis in MCF7 cells. Cells (1 × 10 6 per dish) were plated in 100-mm dishes 24h before treatments. Cells were then treated without or with panobinostat (10nM) for 0-180min (A-C) or other HDAC inhibitors for 45min (D-F). Following treatments, cells were collected, extracted with lysis buffer, and processed for either immunoprecipitation or immunoblotting studies, as described in Materials and methods . Representative immunoblots illustrate acetylation (Ac-StAR) and expression of StAR (T-StAR) in different treatment groups. IgG heavy chain (IgG-Hc) and β-actin expression were assessed for loading controls in immunoprecipitation and immunoblotting, respectively. Integrated optical density (IOD) values of Ac-StAR and T-StAR in each band were quantified and normalized with corresponding IgG-Hc and β-actin expression, and presented as fold response (C,F). E2 levels in media at each time point were determined by ELISA and presented as pg/mg protein (C). Data are representative of 3-4 independent experiments. *, p

    Article Snippet: 17β-estradiol (E2) levels in cell culture media were determined using ELISA Kit from Cayman Chemical (Ann Arbor, MI), according to manufacturer’s protocol [ ], optimized further for better efficacy.

    Techniques: Expressing, Lysis, Immunoprecipitation, Western Blot, Enzyme-linked Immunosorbent Assay

    Relative expression of StAR and aromatase proteins, and E2 synthesis, in human normal mammary epithelial (MCF10A and MCF12F), ER+ breast cancer (MCF7, MBA-MD-361, and T-47D), TNBC (MBA-MD-468, MBA-MD-231, and BT-549), and H295R, cell lines. These cells were cultured with appropriate media, and were harvested in RIPA buffer at 75-80% confluence. Cells and media were then processed for whole cell extract preparation and E2 extraction by diethyl ether, respectively. Representative immunoblots illustrate total StAR (T-StAR) and aromatase protein levels in different cell lines (A). Immunoblots shown are representative of 4 independent experiments. β-actin expression was assessed as a loading control. B, E2 levels in media from different cell lines were determined by ELISA and presented as pg/mg protein (n=4, ±SE). Different letters above the bars indicate that these groups differ significantly from each other at least at p

    Journal: Biochemical and biophysical research communications

    Article Title: Overexpression of the steroidogenic acute regulatory protein in breast cancer: Regulation by histone deacetylase inhibition

    doi: 10.1016/j.bbrc.2018.12.145

    Figure Lengend Snippet: Relative expression of StAR and aromatase proteins, and E2 synthesis, in human normal mammary epithelial (MCF10A and MCF12F), ER+ breast cancer (MCF7, MBA-MD-361, and T-47D), TNBC (MBA-MD-468, MBA-MD-231, and BT-549), and H295R, cell lines. These cells were cultured with appropriate media, and were harvested in RIPA buffer at 75-80% confluence. Cells and media were then processed for whole cell extract preparation and E2 extraction by diethyl ether, respectively. Representative immunoblots illustrate total StAR (T-StAR) and aromatase protein levels in different cell lines (A). Immunoblots shown are representative of 4 independent experiments. β-actin expression was assessed as a loading control. B, E2 levels in media from different cell lines were determined by ELISA and presented as pg/mg protein (n=4, ±SE). Different letters above the bars indicate that these groups differ significantly from each other at least at p

    Article Snippet: 17β-estradiol (E2) levels in cell culture media were determined using ELISA Kit from Cayman Chemical (Ann Arbor, MI), according to manufacturer’s protocol [ ], optimized further for better efficacy.

    Techniques: Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay

    Transforming growth factor-β (TGF-β) induces the colorectal cancer cells metastasis through the epithelial to mesenchymal transition progress. (A) Imagesof CT26 and HCT116 cells treated with phosphate buffered saline (PBS), TGF-β (5 ng/mL), tumor-associated macrophages (TAMs) cultural supernatant and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours. Scale bar=30 μm. (B) Relative E-cadherin expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (C) Relative vimentin expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (D) Western blotting of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for24 hours (E) Immunofluorescence of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (F) Immunofluorescence of E-cadherin andvimentin in tumor tissues of non-metastatic colorectal cancer patients (patients/N) andmetastatic colorectal cancer patients (patients/M). Scalebar=20 μm. The data was presented as the mean±standard error of mean from three independent experiments. *p

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Tumor-Associated Macrophages Derived TGF-β‒Induced Epithelial to Mesenchymal Transition in Colorectal Cancer Cells through Smad2,3-4/Snail Signaling Pathway

    doi: 10.4143/crt.2017.613

    Figure Lengend Snippet: Transforming growth factor-β (TGF-β) induces the colorectal cancer cells metastasis through the epithelial to mesenchymal transition progress. (A) Imagesof CT26 and HCT116 cells treated with phosphate buffered saline (PBS), TGF-β (5 ng/mL), tumor-associated macrophages (TAMs) cultural supernatant and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours. Scale bar=30 μm. (B) Relative E-cadherin expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (C) Relative vimentin expression of CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for 24 hours (n=3). (D) Western blotting of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL) for24 hours (E) Immunofluorescence of E-cadherin and vimentin in CT26 and HCT116 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (F) Immunofluorescence of E-cadherin andvimentin in tumor tissues of non-metastatic colorectal cancer patients (patients/N) andmetastatic colorectal cancer patients (patients/M). Scalebar=20 μm. The data was presented as the mean±standard error of mean from three independent experiments. *p

    Article Snippet: Samples were blocked in 5% bovine serum albumin in phosphate buffered saline (PBS) for 1 hour, E-cadherin (1:200, Abcam) and vimentin (1:300, Abcam) were incubated at 4°C overnight, followed by signal amplification using TSA Kit (PerkinElmer, Waltham, MA) [ ].

    Techniques: Expressing, Western Blot, Immunofluorescence

    Transforming growth factor-β (TGF-β) mediates the colorectal cancer cells epithelial to mesenchymal transition progress via Smad2,3-4/Snail/E-cadherin pathway. (A) Western blotting of p-Smad2, p-Smad3, Smad4, Snail in CT26 cells treated with phosphate buffered saline (PBS), TGF-β (5 ng/mL), tumor-associated macrophages (TAMs) cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). (B) Immunofluorescence of Smad2/3 in CT26 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (C) Immunofluorescence of Smad4 in CT26 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (D) Images of CT26 cells treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=20 μm. (E) Relative migrant cell number in 104 CT26 cells treated with PBS, TGF-β, and TGF-β combined with Snail siRNA (n=3). (F) The wound-healing assay for evaluating the migration of CT26 treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=100 μm. (G) Immunofluorescence of E-cadherin in CT26 treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=10 μm. (H) The lung weight of tumor-bearing mice treated with PBS or LY2109761 (n=9). (I) The pulmonary tumor nodules of tumor-bearing mice treated with PBS or LY2109761 (n=9). (J) H E staining of a representative lung of tumor-bearing mice treated with PBS or LY2109761. Scale bar=200 μm. (K) Schema of TAMs inducing the colorectal cancer cells metastasis. The data was presented as the mean±standard error of mean from three independent experiments. *p

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Tumor-Associated Macrophages Derived TGF-β‒Induced Epithelial to Mesenchymal Transition in Colorectal Cancer Cells through Smad2,3-4/Snail Signaling Pathway

    doi: 10.4143/crt.2017.613

    Figure Lengend Snippet: Transforming growth factor-β (TGF-β) mediates the colorectal cancer cells epithelial to mesenchymal transition progress via Smad2,3-4/Snail/E-cadherin pathway. (A) Western blotting of p-Smad2, p-Smad3, Smad4, Snail in CT26 cells treated with phosphate buffered saline (PBS), TGF-β (5 ng/mL), tumor-associated macrophages (TAMs) cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). (B) Immunofluorescence of Smad2/3 in CT26 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (C) Immunofluorescence of Smad4 in CT26 cells treated with PBS, TGF-β (5 ng/mL), TAMs cultural supernatant, and TAMs cultural supernatant combined with TGF-β neutralizing antibody (10 ng/mL). Scale bar=10 μm. (D) Images of CT26 cells treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=20 μm. (E) Relative migrant cell number in 104 CT26 cells treated with PBS, TGF-β, and TGF-β combined with Snail siRNA (n=3). (F) The wound-healing assay for evaluating the migration of CT26 treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=100 μm. (G) Immunofluorescence of E-cadherin in CT26 treated with PBS, TGF-β, and TGF-β combined with Snail siRNA. Scale bar=10 μm. (H) The lung weight of tumor-bearing mice treated with PBS or LY2109761 (n=9). (I) The pulmonary tumor nodules of tumor-bearing mice treated with PBS or LY2109761 (n=9). (J) H E staining of a representative lung of tumor-bearing mice treated with PBS or LY2109761. Scale bar=200 μm. (K) Schema of TAMs inducing the colorectal cancer cells metastasis. The data was presented as the mean±standard error of mean from three independent experiments. *p

    Article Snippet: Samples were blocked in 5% bovine serum albumin in phosphate buffered saline (PBS) for 1 hour, E-cadherin (1:200, Abcam) and vimentin (1:300, Abcam) were incubated at 4°C overnight, followed by signal amplification using TSA Kit (PerkinElmer, Waltham, MA) [ ].

    Techniques: Western Blot, Immunofluorescence, Wound Healing Assay, Migration, Mouse Assay, Staining

    Antitumor potency of the SA-IL-2 MCSCs vaccine in the pulmonary model mice. a In the cytotoxicity assay, the portion of CTL in the experimental group was significantly higher than in the four control groups. b In ELISA assay, the expression of serum IgG antibodies in the experimental group was significantly higher than in the four control groups. c FCM analysis showed that the portion of DCs (CD11c + CD80 + ) in the experimental group was significantly larger than in the four control groups. d FCM analysis showed that the portion of CD8 + and CD4 + T cells in the experimental group was significantly larger than in the four control groups. *P

    Journal: Stem Cell Research & Therapy

    Article Title: Development of a therapy against metastatic bladder cancer using an interleukin-2 surface-modified MB49 bladder cancer stem cells vaccine

    doi: 10.1186/s13287-015-0211-1

    Figure Lengend Snippet: Antitumor potency of the SA-IL-2 MCSCs vaccine in the pulmonary model mice. a In the cytotoxicity assay, the portion of CTL in the experimental group was significantly higher than in the four control groups. b In ELISA assay, the expression of serum IgG antibodies in the experimental group was significantly higher than in the four control groups. c FCM analysis showed that the portion of DCs (CD11c + CD80 + ) in the experimental group was significantly larger than in the four control groups. d FCM analysis showed that the portion of CD8 + and CD4 + T cells in the experimental group was significantly larger than in the four control groups. *P

    Article Snippet: The concentrations of IgG were measured using ELISA kits (Abcam) according to the manufacturer’s protocol.

    Techniques: Mouse Assay, Cytotoxicity Assay, CTL Assay, Enzyme-linked Immunosorbent Assay, Expressing

    Antitumor potency of the SA-IL-2 MCSCs vaccine in the subcutaneous model mice. a In the cytotoxicity assay, the portion of CTL in the experimental group was significantly higher than in the four control groups. b In ELISA assay, the expression of serum IgG antibodies in the experimental group was significantly higher than in the four control groups. c FCM analysis showed that the portion of DCs (CD11c + CD80 + ) in the experimental group was significantly larger than in the four control groups. d FCM analysis showed that the portion of CD8 + and CD4 + T cells in the experimental group was significantly larger than in the four control groups. *P

    Journal: Stem Cell Research & Therapy

    Article Title: Development of a therapy against metastatic bladder cancer using an interleukin-2 surface-modified MB49 bladder cancer stem cells vaccine

    doi: 10.1186/s13287-015-0211-1

    Figure Lengend Snippet: Antitumor potency of the SA-IL-2 MCSCs vaccine in the subcutaneous model mice. a In the cytotoxicity assay, the portion of CTL in the experimental group was significantly higher than in the four control groups. b In ELISA assay, the expression of serum IgG antibodies in the experimental group was significantly higher than in the four control groups. c FCM analysis showed that the portion of DCs (CD11c + CD80 + ) in the experimental group was significantly larger than in the four control groups. d FCM analysis showed that the portion of CD8 + and CD4 + T cells in the experimental group was significantly larger than in the four control groups. *P

    Article Snippet: The concentrations of IgG were measured using ELISA kits (Abcam) according to the manufacturer’s protocol.

    Techniques: Mouse Assay, Cytotoxicity Assay, CTL Assay, Enzyme-linked Immunosorbent Assay, Expressing

    Impaired insulin secretion from isolated Pick1 cKO islets. (a) GSIS was performed on isolated islets stimulated with 4.8 or 30 mM KCl or 2.8 or 16.7 mM glucose (Glc). (b) The stimulated index was also calculated following KCl or Glc treatment. (c) Insulin content in the isolated islets was measured by ELISA. (d) Isolated WT and Pick1 cKO islets were incubated in the presence of 16.7 mM Glc at 2-min intervals for 20 min. (e) Same data as shown in d, but plotted as release per 2 min. (f, g) Secreted insulin was tested from isolated WT and Pick1 cKO islets incubated with or without E4. Data are represented as mean ± SEM, n = 3, three to four groups per type of mouse, 10–0 islets per group. * p

    Journal: Molecular Biology of the Cell

    Article Title: PICK1 is essential for insulin production and the maintenance of glucose homeostasis

    doi: 10.1091/mbc.E17-03-0204

    Figure Lengend Snippet: Impaired insulin secretion from isolated Pick1 cKO islets. (a) GSIS was performed on isolated islets stimulated with 4.8 or 30 mM KCl or 2.8 or 16.7 mM glucose (Glc). (b) The stimulated index was also calculated following KCl or Glc treatment. (c) Insulin content in the isolated islets was measured by ELISA. (d) Isolated WT and Pick1 cKO islets were incubated in the presence of 16.7 mM Glc at 2-min intervals for 20 min. (e) Same data as shown in d, but plotted as release per 2 min. (f, g) Secreted insulin was tested from isolated WT and Pick1 cKO islets incubated with or without E4. Data are represented as mean ± SEM, n = 3, three to four groups per type of mouse, 10–0 islets per group. * p

    Article Snippet: The serum insulin and proinsulin and the insulin from the medium and islet supernatants collected from the GSIS were tested using the Ultra-sensitive mouse insulin ELISA Kit (Crystal Chem) or the Proinsulin rat/mouse ELISA Kit (Mercodia Insulin/Proinsulin).

    Techniques: Isolation, Gas Chromatography, Enzyme-linked Immunosorbent Assay, Incubation

    Characterization of IFN-γ ( a , e ), IL-6 ( b , f ), TNF-α ( c , g ) and Lptn ( d , f ) secretion from the spleen ( a , b , c and d ) and lung ( a , b , c and d ) tissues of mice after CIV alone, S. pseudintermedius alone, or both. The concentrations in the supernatants of homogenates from the spleens and lungs on days 1, 3 and 5 post-challenge were measured by ELISA. The results are expressed in terms of pg/ml. Analysis of variance (ANOVA) was performed for the statistical analysis. *, P

    Journal: BMC Veterinary Research

    Article Title: Canine influenza virus coinfection with Staphylococcus pseudintermedius enhances bacterial colonization, virus load and clinical presentation in mice

    doi: 10.1186/s12917-016-0708-6

    Figure Lengend Snippet: Characterization of IFN-γ ( a , e ), IL-6 ( b , f ), TNF-α ( c , g ) and Lptn ( d , f ) secretion from the spleen ( a , b , c and d ) and lung ( a , b , c and d ) tissues of mice after CIV alone, S. pseudintermedius alone, or both. The concentrations in the supernatants of homogenates from the spleens and lungs on days 1, 3 and 5 post-challenge were measured by ELISA. The results are expressed in terms of pg/ml. Analysis of variance (ANOVA) was performed for the statistical analysis. *, P

    Article Snippet: ELISAs for the levels of interferon-gamma (IFN-γ), interleukin 6 (IL-6), lymphotactin (Lptn) and tumor necrosis factor (TNF-α) on the lung and spleen from infected mice were performed using ELISA kits (Sigma-Aldrich, Beijing) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay