il-6 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher gene exp il6 mm00446190 m1
    Brain mRNA expression of proinflammatory cytokines, but not sickness behavior, is increased by intracerebroventricular lipopolysaccharide. Both wild-type (WT) and indoleamine-2,3-dioxygenase (IDO1) knockout (KO) mice exhibit increased steady-state mRNA expression of ( a ) IL-1β and ( b ) TNFα, but not of ( c ) <t>IL-6</t> in the whole brain 24 hours post intracerebroventricular (ICV) lipopolysaccharide (LPS, 10 ng). Exploratory locomotor activity measured by ( d ) distance traveled or ( e ) duration of time during the test spent moving were not different 24 hours after ICV LPS. Data are average ± standard error of the mean. n = 4 to 5 mice per group.
    Gene Exp Il6 Mm00446190 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3359 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp il6 mm00446190 m1/product/Thermo Fisher
    Average 99 stars, based on 3359 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 mm00446190 m1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    R&D Systems human il 6
    JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) <t>IL-6</t> and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P
    Human Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human il 6/product/R&D Systems
    Average 99 stars, based on 1161 article reviews
    Price from $9.99 to $1999.99
    human il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp il6 hs00985639 m1
    Cytokine content of tissue culture supernatants Culture supernatants arising from 20 IRIS and 19 non-IRIS controls were assayed by Luminex. IL-4 and IL-17, whose levels were close to lower detection limits, did not differ between TB-IRIS and non-IRIS. Otherwise levels were consistently and significantly higher in TB-IRIS. After correction of p-values for multiple comparisons the largest and significant fold differences were in IL-12p40, IL-1β, GM-CSF, TNF, IL-10, <t>IL-6,</t> IL-2 and IL-8. p values are uncorrected in this figure with significant associations shown in red
    Gene Exp Il6 Hs00985639 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp il6 hs00985639 m1/product/Thermo Fisher
    Average 99 stars, based on 1168 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 hs00985639 m1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp il6 hs00174131 m1
    Baseline pro-inflammatory monocyte frequency correlation (PB) with disease activity scores in SpA and ERA patients. Correlation in ERA patients: a . baseline TNF% vs JSpADA b . baseline <t>IL-6%</t> vs JspADA. Correlation in SpA patients: c . baseline TNF% vs BASDAI, d . baseline IL-6% vs BASDAI. PB: peripheral blood, BASDAI Bath Ankylosing Spondylitis Disease Activity Index, JspADA Juvenile spondyloarthritis Disease Activity Score
    Gene Exp Il6 Hs00174131 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 903 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp il6 hs00174131 m1/product/Thermo Fisher
    Average 99 stars, based on 903 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 hs00174131 m1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    R&D Systems mouse il 6
    <t>IL-6</t> is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P
    Mouse Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 760 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse il 6/product/R&D Systems
    Average 99 stars, based on 760 article reviews
    Price from $9.99 to $1999.99
    mouse il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    R&D Systems recombinant human il 6
    MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; <t>IL-6,</t> interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.
    Recombinant Human Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 996 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human il 6/product/R&D Systems
    Average 99 stars, based on 996 article reviews
    Price from $9.99 to $1999.99
    recombinant human il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    94
    PeproTech recombinant mouse il 6
    WT-CCN1, but not DM-CCN1, induces <t>IL-6</t> in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.
    Recombinant Mouse Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 437 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mouse il 6/product/PeproTech
    Average 94 stars, based on 437 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 6 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Thermo Fisher mouse il 6
    Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c <t>IL-6</t> and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P
    Mouse Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 553 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse il 6/product/Thermo Fisher
    Average 99 stars, based on 553 article reviews
    Price from $9.99 to $1999.99
    mouse il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Abcam 6 well plates
    Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a <t>6-well</t> plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.
    6 Well Plates, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/6 well plates/product/Abcam
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    6 well plates - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Abcam anti il 6
    Comparison of TNF- α and <t>IL-6</t> production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p
    Anti Il 6, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il 6/product/Abcam
    Average 92 stars, based on 729 article reviews
    Price from $9.99 to $1999.99
    anti il 6 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Thermo Fisher human il 6
    STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human <t>IL-6</t> for 30 min. Whole-cell lysates
    Human Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human il 6/product/Thermo Fisher
    Average 93 stars, based on 580 article reviews
    Price from $9.99 to $1999.99
    human il 6 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    99
    PeproTech recombinant human il 6
    Anti-IL-6R reduces the expression of IFN-γ in activated Xiap −/− Treg cells. a , b Anti-IL-6R decreases IFN-γ expression in re-stimulated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated with anti-CD3/CD28 and IL-2, or with additional IL-12 as indicated, in the absence or presence of anti-IL-6R (50 μg ml −1 ) for 4 days. iTreg cells were re-stimulated with TPA/A23187 for 24 h and IFN-γ production was determined by ELISA ( a ), or reactivated with TPA/A23187 for 5 h and the expressions of Foxp3 and IFN-γ were determined by intracellular staining ( b ). c , d Anti-IL-6R inhibits IFN-γ expression in human Treg cells. Control and human XIAP-knockdown iTreg cells were stimulated as in ( a , b ), with additional <t>IL-6</t> as indicated, and secretion ( c ) or intracellular expression ( d ) of IFN-γ was determined. e Inability of anti-TNF or anti-IL-1R to inhibit the production of IFN-γ in activated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated, as described in ( a ), in the presence or absence of anti-TNF or anti-IL-1R (50 μg ml −1 each) for 4 days. Treg cells were re-stimulated with TPA/A23187 for 24 h and the secreted IFN-γ was determined by ELISA. f Anti-IL-6R rescues the impaired suppressive activity of Xiap −/− tTreg cells in vivo. CD45.2 + WT or Xiap −/− tTreg cells were co-transferred with CD45.1 + CD4 + CD25 - effector T cells into male CD45.1 + RagI −/− mice. Anti-IL-6R antibody (500 μg per mouse) was intraperitoneally administrated at day 0, followed by weekly dosing of 500 μg. The body weights of mice were monitored at the indicated time-points. *** P
    Recombinant Human Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 661 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human il 6/product/PeproTech
    Average 99 stars, based on 661 article reviews
    Price from $9.99 to $1999.99
    recombinant human il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp il6 rn01410330 m1
    The LPD prevents renal injuries, the impairment of autophagy and the activation of mTORC1 in the kidneys of Wistar fatty rats. ( a ) Representative microphotographs of Masson’s trichrome (MT) staining of glomeruli (scale bar: 100 μm) and the tubulo-interstitial area (scale bar: 1 mm) after the 24-week dietary intervention. ( b ) Glomerular fibrotic score ( n = 3) and ( c ) tubulo-interstitial fibrotic score ( n = 3), assessed using MT staining after the 24-week dietary intervention. mRNA expression of Col3 ( d ), Kim-1 ( e ), Cd68 ( f ), Ccl12 ( g ), Tlr4 ( h ) and <t>Il6</t> ( i ) in the renal cortex, adjusted to 18 s levels, after the 24-week dietary intervention ( n = 6 each). ( j ) Representative photographs of immunoblotting for p62, β-actin, p-S6RP and S6RP in the renal cortex after the 24-week dietary intervention ( n = 3). Quantitative ratios of p62 to β-actin ( k ) and p-S6RP to S6RP ( l ) ( n = 3). Col3: type 3 collagen, Kim-1: kidney injury molecule-1, Ccl2: C-C motif chemokine ligand 2, Tlr4: Toll-like receptor 4, Il6: interleukin-6, p-S6RP: phospho-S6 ribosomal protein, S6RP: S6 ribosomal protein. The data are shown as the mean ± SD. * p
    Gene Exp Il6 Rn01410330 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 550 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp il6 rn01410330 m1/product/Thermo Fisher
    Average 99 stars, based on 550 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 rn01410330 m1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    R&D Systems rmil 6
    IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of <t>rmIL-6.</t> The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p
    Rmil 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 205 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rmil 6/product/R&D Systems
    Average 99 stars, based on 205 article reviews
    Price from $9.99 to $1999.99
    rmil 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Abcam anti il6 antibody
    Pancreatic cancer-educated macrophages induced the upregulation of CD59 in cancer cells via the IL-6R/STAT3 pathway. a Gene expression profiling in AsPC-1 cells cocultured with THP-1 macrophages compared with AsPC-1 cells. b , c GO and KEGG pathway analyses of differentially expressed genes. d <t>IL6-R</t> was extremely elevated in the cocultured group compared with that in the control group. e AsPC-1 and BxPC-3 cells were cultured with THP-1 macrophages and analyzed for the level of total STAT3 or phosphorylated STAT3 (p-STAT3). f , g IL-6 expression in macrophages cocultured with AsPC-1 and BxPC-3 cells was detected by qRT-PCR, western blot and ELISA. h The CD59 expression and phosphorylation of STAT3 in pancreatic cancer cells incubated with various concentrations of recombinant IL-6 (0, 0.01, 1, 10, and 100 nM) were detected by western blot
    Anti Il6 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 270 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il6 antibody/product/Abcam
    Average 99 stars, based on 270 article reviews
    Price from $9.99 to $1999.99
    anti il6 antibody - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Becton Dickinson mouse il 6
    <t>IL-6</t> induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).
    Mouse Il 6, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 334 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse il 6/product/Becton Dickinson
    Average 92 stars, based on 334 article reviews
    Price from $9.99 to $1999.99
    mouse il 6 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    R&D Systems recombinant il 6
    MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, <t>IL-6,</t> TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P
    Recombinant Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 470 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant il 6/product/R&D Systems
    Average 99 stars, based on 470 article reviews
    Price from $9.99 to $1999.99
    recombinant il 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Becton Dickinson interleukin il 6
    Influence of erythropoietin on meningococcal-induced cytokine production. <t>Interleukin</t> <t>(IL)-6</t> and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as
    Interleukin Il 6, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 364 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/interleukin il 6/product/Becton Dickinson
    Average 92 stars, based on 364 article reviews
    Price from $9.99 to $1999.99
    interleukin il 6 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher il 6 mouse elisa kit
    Determination of levels of TNF-α and <t>IL-6</t> in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial <t>ELISA</t> kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).
    Il 6 Mouse Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 6 mouse elisa kit/product/Thermo Fisher
    Average 99 stars, based on 222 article reviews
    Price from $9.99 to $1999.99
    il 6 mouse elisa kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Abcam interleukin 6
    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic <t>IL-6</t> levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P
    Interleukin 6, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/interleukin 6/product/Abcam
    Average 99 stars, based on 352 article reviews
    Price from $9.99 to $1999.99
    interleukin 6 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology anti il 6
    Western blot analysis of protein expression of <t>IL-6,</t> p-STAT3, survivin, STAT3, and VEGF. Protein levels of IL-6, Survivin, p-STAT3, STAT3, and VEGF in normal gastric and tumor tissue were determined using western blotting. Beta-actin was a loading control. Relative protein expression of IL-6 (A), VEGF (B), surviving (C), p-STAT3 (D) was normalized to of the corresponding beta-actin level. Positive immunoreactive bands were quantified densitometrically and expressed as IL-6, Survivin, p-STAT3, STAT3, and VEGF in optical density units, respectively. * P
    Anti Il 6, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 344 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il 6/product/Santa Cruz Biotechnology
    Average 93 stars, based on 344 article reviews
    Price from $9.99 to $1999.99
    anti il 6 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    94
    R&D Systems il6
    Sorafenib regulates the expression of <t>ID1/p16/IL6.</t> a RT-PCR was conducted in HepG2 cells treated with different concentrations of sorafenib for 24 h. b Western blot experiments were performed in HepG2 cells treated with different concentrations of sorafenib for 24 h. c Cell supernatant was collected to perform the IL6 ELISA assay. d After incubated with 5 μM sorafenib for 24 h, cells were stained with SA-β-gal. * p
    Il6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 507 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il6/product/R&D Systems
    Average 94 stars, based on 507 article reviews
    Price from $9.99 to $1999.99
    il6 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp il6 mm99999064 m1
    Reduced inflammatory cytokine expression in retina from Lodamin-administered mice, and summary of Lodamin’s activity on T cells. (A) On day 21 after IRBP immunization, retinas were harvested and total RNA was extracted from naive mice (unimmunized control), EAU control group and Lodamin-administered group. The expression of <t>IL-6,</t> TNF, IFN-γ, and IL-17A were analyzed by realtime qPCR (mean ± SEM, n = 5–8, *** p
    Gene Exp Il6 Mm99999064 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 262 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp il6 mm99999064 m1/product/Thermo Fisher
    Average 99 stars, based on 262 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 mm99999064 m1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Brain mRNA expression of proinflammatory cytokines, but not sickness behavior, is increased by intracerebroventricular lipopolysaccharide. Both wild-type (WT) and indoleamine-2,3-dioxygenase (IDO1) knockout (KO) mice exhibit increased steady-state mRNA expression of ( a ) IL-1β and ( b ) TNFα, but not of ( c ) IL-6 in the whole brain 24 hours post intracerebroventricular (ICV) lipopolysaccharide (LPS, 10 ng). Exploratory locomotor activity measured by ( d ) distance traveled or ( e ) duration of time during the test spent moving were not different 24 hours after ICV LPS. Data are average ± standard error of the mean. n = 4 to 5 mice per group.

    Journal: Journal of Neuroinflammation

    Article Title: Intracerebroventricular administration of lipopolysaccharide induces indoleamine-2,3-dioxygenase-dependent depression-like behaviors

    doi: 10.1186/1742-2094-10-87

    Figure Lengend Snippet: Brain mRNA expression of proinflammatory cytokines, but not sickness behavior, is increased by intracerebroventricular lipopolysaccharide. Both wild-type (WT) and indoleamine-2,3-dioxygenase (IDO1) knockout (KO) mice exhibit increased steady-state mRNA expression of ( a ) IL-1β and ( b ) TNFα, but not of ( c ) IL-6 in the whole brain 24 hours post intracerebroventricular (ICV) lipopolysaccharide (LPS, 10 ng). Exploratory locomotor activity measured by ( d ) distance traveled or ( e ) duration of time during the test spent moving were not different 24 hours after ICV LPS. Data are average ± standard error of the mean. n = 4 to 5 mice per group.

    Article Snippet: Real-time RT-PCR was performed over 40 cycles using a CFX384™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) and prevalidated Taqman® Gene Expression Assays (Life Technologies) for both target and housekeeping control genes: GAPDH (Mm99999915_g1), IL-1β (Mm01336189_m1), TNFα (Mm00443258_m1), and IL-6 (Mm00446190_m1).

    Techniques: Expressing, Knock-Out, Mouse Assay, Activity Assay

    JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) IL-6 and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) IL-6 and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Micro-CT

    Effects of JAK2 and STAT3 on ATII to mesenchymal and fibroblast to myofibroblast transitions. Primary ATII and lung fibroblasts were isolated from the lungs of IPF patients. a The cells were incubated with the dual p-JAK2/p-STAT3 inhibitor JSI-124 for 30 min followed by TGF-β1 stimulation for an additional 24 h. IL-6 and IL-13 levels in cell supernatants were measured using ELISA. b Ratios of JAK2/p-JAK2 and STAT3/p-STAT3 were determined by western blotting in ATII cells stimulated for 40 min or 24 h with TGF-β1 in the presence or absence of JSI-124. c , d ATII cells were pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124, and then stimulated for 72 h with TGF-β1 ( c ) or IL-6/IL-13 ( d ). e Levels of IL-6 and IL-13 in primary fibroblasts. f JAK2/p-JAK2 and STAT3/p-STAT3 protein expression in human lung fibroblasts. g , h Primary lung fibroblasts pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and stimulated for 72 h with TGF-β1 ( g ) or IL-6/IL-13 ( h ). Representative western blots are shown. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Effects of JAK2 and STAT3 on ATII to mesenchymal and fibroblast to myofibroblast transitions. Primary ATII and lung fibroblasts were isolated from the lungs of IPF patients. a The cells were incubated with the dual p-JAK2/p-STAT3 inhibitor JSI-124 for 30 min followed by TGF-β1 stimulation for an additional 24 h. IL-6 and IL-13 levels in cell supernatants were measured using ELISA. b Ratios of JAK2/p-JAK2 and STAT3/p-STAT3 were determined by western blotting in ATII cells stimulated for 40 min or 24 h with TGF-β1 in the presence or absence of JSI-124. c , d ATII cells were pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124, and then stimulated for 72 h with TGF-β1 ( c ) or IL-6/IL-13 ( d ). e Levels of IL-6 and IL-13 in primary fibroblasts. f JAK2/p-JAK2 and STAT3/p-STAT3 protein expression in human lung fibroblasts. g , h Primary lung fibroblasts pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and stimulated for 72 h with TGF-β1 ( g ) or IL-6/IL-13 ( h ). Representative western blots are shown. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

    Fibroblast migration and proliferation in IPF were dependent on JAK2 and STAT3 activation. a , b Primary human fibroblasts from IPF patients were pre-treated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and then cell migration was assessed. A scrape-wound was created by using a sterile p200 pipette tip to make a perpendicular linear scratch in the culture. After the cells had been washed, culture medium with or without pharmacologic modulators and IL-6/IL-13 was added. The size of the wound remaining was analyzed at the indicated times and expressed as a percentage of the initial wound area. c Fibroblast proliferation during 48 h was evaluated by the BrdU assay. Different JAK2 and STAT3 inhibitors were added 30 min before 10% fetal bovine serum (FBS) or ( c ) 50 ng IL-6/IL-13/mL ( d ) was added as the stimulus. Values are expressed as relative absorbance (450 nm) units. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Fibroblast migration and proliferation in IPF were dependent on JAK2 and STAT3 activation. a , b Primary human fibroblasts from IPF patients were pre-treated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and then cell migration was assessed. A scrape-wound was created by using a sterile p200 pipette tip to make a perpendicular linear scratch in the culture. After the cells had been washed, culture medium with or without pharmacologic modulators and IL-6/IL-13 was added. The size of the wound remaining was analyzed at the indicated times and expressed as a percentage of the initial wound area. c Fibroblast proliferation during 48 h was evaluated by the BrdU assay. Different JAK2 and STAT3 inhibitors were added 30 min before 10% fetal bovine serum (FBS) or ( c ) 50 ng IL-6/IL-13/mL ( d ) was added as the stimulus. Values are expressed as relative absorbance (450 nm) units. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Migration, Activation Assay, Transferring, BrdU Staining

    Dual JAK2/STAT3 gene silencing suppressed ATII to mesenchymal and fibroblast to myofibroblast transitions. ATII A549 and human lung fibroblast MRC5 cell lines were transfected with control siRNA(−), siRNA-JAK2, siRNA-STAT3, or both siRNA-JAK2/STAT3 and stimulated for 72 h with TG-Fβ1 or IL-6/IL-13 at a concentration of 5 ng/mL. a Total protein and RNA from cell lysates were analyzed by ( a , b ) western blot and ( c ) qPCR. Mesenchymal collagen type I and epithelial E-cadherin markers were measured using ( a ) western blot and ( c ) qPCR. Senescence p21, autophagy LC3I/II, and anti-apoptotic BCL-2 markers were ( a ) measured using western blot and ( b ) quantified by densitometry. Data are expressed as the ratio to β-actin protein and mRNA levels. The results are expressed as the mean (SEM) of n = 4 independent experiments per condition. One-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Dual JAK2/STAT3 gene silencing suppressed ATII to mesenchymal and fibroblast to myofibroblast transitions. ATII A549 and human lung fibroblast MRC5 cell lines were transfected with control siRNA(−), siRNA-JAK2, siRNA-STAT3, or both siRNA-JAK2/STAT3 and stimulated for 72 h with TG-Fβ1 or IL-6/IL-13 at a concentration of 5 ng/mL. a Total protein and RNA from cell lysates were analyzed by ( a , b ) western blot and ( c ) qPCR. Mesenchymal collagen type I and epithelial E-cadherin markers were measured using ( a ) western blot and ( c ) qPCR. Senescence p21, autophagy LC3I/II, and anti-apoptotic BCL-2 markers were ( a ) measured using western blot and ( b ) quantified by densitometry. Data are expressed as the ratio to β-actin protein and mRNA levels. The results are expressed as the mean (SEM) of n = 4 independent experiments per condition. One-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Transfection, Concentration Assay, Western Blot, Real-time Polymerase Chain Reaction

    Cytokine content of tissue culture supernatants Culture supernatants arising from 20 IRIS and 19 non-IRIS controls were assayed by Luminex. IL-4 and IL-17, whose levels were close to lower detection limits, did not differ between TB-IRIS and non-IRIS. Otherwise levels were consistently and significantly higher in TB-IRIS. After correction of p-values for multiple comparisons the largest and significant fold differences were in IL-12p40, IL-1β, GM-CSF, TNF, IL-10, IL-6, IL-2 and IL-8. p values are uncorrected in this figure with significant associations shown in red

    Journal: The European respiratory journal

    Article Title: Hypercytokinaemia accompanies HIV-tuberculosis immune reconstitution inflammatory syndrome

    doi: 10.1183/09031936.00091010

    Figure Lengend Snippet: Cytokine content of tissue culture supernatants Culture supernatants arising from 20 IRIS and 19 non-IRIS controls were assayed by Luminex. IL-4 and IL-17, whose levels were close to lower detection limits, did not differ between TB-IRIS and non-IRIS. Otherwise levels were consistently and significantly higher in TB-IRIS. After correction of p-values for multiple comparisons the largest and significant fold differences were in IL-12p40, IL-1β, GM-CSF, TNF, IL-10, IL-6, IL-2 and IL-8. p values are uncorrected in this figure with significant associations shown in red

    Article Snippet: We used the following TaqMan® Gene Expression Assays: IL-1β, Catalogue Number Hs00174097_m1; IL-2, Hs00174114_m1; IL-4, Hs00174122_m1; IL-5, Hs00174200_m1; IL-6, Hs00985639_m1; IFN-γ, Hs00174143_m1; TNF, Hs00174128_m1; IL-8, Hs01038788_m1; IL-10, Hs00174086_m1; IL-12p40, Hs01011518_m1; lL-13, Hs00174379_m1; IL-15, Hs00542562_m1; IL-17A, Hs00174383_m1; IL-18, Hs01038788_m1; GM-CSF, Hs00171266_m1; TGF-β1, Hs00171257_m1.

    Techniques: Luminex

    Average log fold induction of cytokine genes by heat killed M. tuberculosis in TB-IRIS and non-IRIS patients PBMC from 22 TB-IRIS and 22 non-IRIS control patients were cultured in the presence or absence of heat killed M. tuberculosis H37Rv for 6 and 24 hours. At the end of the culture period the cells were lysed, RNA extracted and used in quantitative RT-PCR. The fold induction of genes was calculated by the ΔΔCt method and values normalised by log 10 transformation. At 6 hours, induction was significantly higher in TB-IRIS than non-IRIS for IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, IL-15, IL-17A, IFN-γ, GM-CSF and TNF (p ≤ 0.05). At 24 hours significant differences between TB-IRIS and non-IRIS were present for IL-8, IL-10, IL-15 (higher in non-IRIS) and TGF-β1. p values are uncorrected in this figure with significant associations shown in red

    Journal: The European respiratory journal

    Article Title: Hypercytokinaemia accompanies HIV-tuberculosis immune reconstitution inflammatory syndrome

    doi: 10.1183/09031936.00091010

    Figure Lengend Snippet: Average log fold induction of cytokine genes by heat killed M. tuberculosis in TB-IRIS and non-IRIS patients PBMC from 22 TB-IRIS and 22 non-IRIS control patients were cultured in the presence or absence of heat killed M. tuberculosis H37Rv for 6 and 24 hours. At the end of the culture period the cells were lysed, RNA extracted and used in quantitative RT-PCR. The fold induction of genes was calculated by the ΔΔCt method and values normalised by log 10 transformation. At 6 hours, induction was significantly higher in TB-IRIS than non-IRIS for IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, IL-15, IL-17A, IFN-γ, GM-CSF and TNF (p ≤ 0.05). At 24 hours significant differences between TB-IRIS and non-IRIS were present for IL-8, IL-10, IL-15 (higher in non-IRIS) and TGF-β1. p values are uncorrected in this figure with significant associations shown in red

    Article Snippet: We used the following TaqMan® Gene Expression Assays: IL-1β, Catalogue Number Hs00174097_m1; IL-2, Hs00174114_m1; IL-4, Hs00174122_m1; IL-5, Hs00174200_m1; IL-6, Hs00985639_m1; IFN-γ, Hs00174143_m1; TNF, Hs00174128_m1; IL-8, Hs01038788_m1; IL-10, Hs00174086_m1; IL-12p40, Hs01011518_m1; lL-13, Hs00174379_m1; IL-15, Hs00542562_m1; IL-17A, Hs00174383_m1; IL-18, Hs01038788_m1; GM-CSF, Hs00171266_m1; TGF-β1, Hs00171257_m1.

    Techniques: Cell Culture, Quantitative RT-PCR, Transformation Assay

    Effect of steroid therapy on cytokines in serum samples Serum concentrations of IFN-γ, IL-6 and TNF were determined in a subset of 10 TB-IRIS who received corticosteroid therapy for 4 weeks. The concentrations of IL-6 and TNF significantly declined whereas no effect on IFN-γ concentrations was observed.

    Journal: The European respiratory journal

    Article Title: Hypercytokinaemia accompanies HIV-tuberculosis immune reconstitution inflammatory syndrome

    doi: 10.1183/09031936.00091010

    Figure Lengend Snippet: Effect of steroid therapy on cytokines in serum samples Serum concentrations of IFN-γ, IL-6 and TNF were determined in a subset of 10 TB-IRIS who received corticosteroid therapy for 4 weeks. The concentrations of IL-6 and TNF significantly declined whereas no effect on IFN-γ concentrations was observed.

    Article Snippet: We used the following TaqMan® Gene Expression Assays: IL-1β, Catalogue Number Hs00174097_m1; IL-2, Hs00174114_m1; IL-4, Hs00174122_m1; IL-5, Hs00174200_m1; IL-6, Hs00985639_m1; IFN-γ, Hs00174143_m1; TNF, Hs00174128_m1; IL-8, Hs01038788_m1; IL-10, Hs00174086_m1; IL-12p40, Hs01011518_m1; lL-13, Hs00174379_m1; IL-15, Hs00542562_m1; IL-17A, Hs00174383_m1; IL-18, Hs01038788_m1; GM-CSF, Hs00171266_m1; TGF-β1, Hs00171257_m1.

    Techniques:

    Serum cytokine concentrations Luminex assays of the serum concentration of 19 IRIS and 20-non-IRIS controls for the most consistently discriminatory cytokines taken from the same patients at the same time. After correction of p-values for multiple comparisons the largest and significant fold differences were in TNF, IL-6, and IFN-γ. Four TB-IRIS patients (shown in grey circles) were classified as having localized (usually lymphadenopathic) disease. In these patient a clear trend towards lower serum cytokine concentrations was seen with 22/24 (92%, 95% CL 74-97%) cytokine values falling on or below the median. p values are uncorrected in this figure.

    Journal: The European respiratory journal

    Article Title: Hypercytokinaemia accompanies HIV-tuberculosis immune reconstitution inflammatory syndrome

    doi: 10.1183/09031936.00091010

    Figure Lengend Snippet: Serum cytokine concentrations Luminex assays of the serum concentration of 19 IRIS and 20-non-IRIS controls for the most consistently discriminatory cytokines taken from the same patients at the same time. After correction of p-values for multiple comparisons the largest and significant fold differences were in TNF, IL-6, and IFN-γ. Four TB-IRIS patients (shown in grey circles) were classified as having localized (usually lymphadenopathic) disease. In these patient a clear trend towards lower serum cytokine concentrations was seen with 22/24 (92%, 95% CL 74-97%) cytokine values falling on or below the median. p values are uncorrected in this figure.

    Article Snippet: We used the following TaqMan® Gene Expression Assays: IL-1β, Catalogue Number Hs00174097_m1; IL-2, Hs00174114_m1; IL-4, Hs00174122_m1; IL-5, Hs00174200_m1; IL-6, Hs00985639_m1; IFN-γ, Hs00174143_m1; TNF, Hs00174128_m1; IL-8, Hs01038788_m1; IL-10, Hs00174086_m1; IL-12p40, Hs01011518_m1; lL-13, Hs00174379_m1; IL-15, Hs00542562_m1; IL-17A, Hs00174383_m1; IL-18, Hs01038788_m1; GM-CSF, Hs00171266_m1; TGF-β1, Hs00171257_m1.

    Techniques: Luminex, Concentration Assay

    Summary of results and implication of TNF and IL-6 in the pathogenesis of TB-IRIS Whilst many pro- and anti-inflammatory cytokine transcript and protein levels were elevated in TB-IRIS patients according to experimental circumstances only IL-6 and TNF were elevated in all circumstances. Thus blockade of IL-6 or TNF may be a rational approach to immunomodulation in this condition.

    Journal: The European respiratory journal

    Article Title: Hypercytokinaemia accompanies HIV-tuberculosis immune reconstitution inflammatory syndrome

    doi: 10.1183/09031936.00091010

    Figure Lengend Snippet: Summary of results and implication of TNF and IL-6 in the pathogenesis of TB-IRIS Whilst many pro- and anti-inflammatory cytokine transcript and protein levels were elevated in TB-IRIS patients according to experimental circumstances only IL-6 and TNF were elevated in all circumstances. Thus blockade of IL-6 or TNF may be a rational approach to immunomodulation in this condition.

    Article Snippet: We used the following TaqMan® Gene Expression Assays: IL-1β, Catalogue Number Hs00174097_m1; IL-2, Hs00174114_m1; IL-4, Hs00174122_m1; IL-5, Hs00174200_m1; IL-6, Hs00985639_m1; IFN-γ, Hs00174143_m1; TNF, Hs00174128_m1; IL-8, Hs01038788_m1; IL-10, Hs00174086_m1; IL-12p40, Hs01011518_m1; lL-13, Hs00174379_m1; IL-15, Hs00542562_m1; IL-17A, Hs00174383_m1; IL-18, Hs01038788_m1; GM-CSF, Hs00171266_m1; TGF-β1, Hs00171257_m1.

    Techniques:

    Baseline pro-inflammatory monocyte frequency correlation (PB) with disease activity scores in SpA and ERA patients. Correlation in ERA patients: a . baseline TNF% vs JSpADA b . baseline IL-6% vs JspADA. Correlation in SpA patients: c . baseline TNF% vs BASDAI, d . baseline IL-6% vs BASDAI. PB: peripheral blood, BASDAI Bath Ankylosing Spondylitis Disease Activity Index, JspADA Juvenile spondyloarthritis Disease Activity Score

    Journal: Pediatric Rheumatology Online Journal

    Article Title: Patients with enthesitis related arthritis show similar monocyte function pattern as seen in adult axial spondyloarthropathy

    doi: 10.1186/s12969-020-0403-9

    Figure Lengend Snippet: Baseline pro-inflammatory monocyte frequency correlation (PB) with disease activity scores in SpA and ERA patients. Correlation in ERA patients: a . baseline TNF% vs JSpADA b . baseline IL-6% vs JspADA. Correlation in SpA patients: c . baseline TNF% vs BASDAI, d . baseline IL-6% vs BASDAI. PB: peripheral blood, BASDAI Bath Ankylosing Spondylitis Disease Activity Index, JspADA Juvenile spondyloarthritis Disease Activity Score

    Article Snippet: Taqman gene expression assay kits were purchased from Applied Biosystems (CA, USA), the IDs being Hs00174128_m1 (TNF), Hs00174131_m1 (IL-6) and HS02786624_g1 (GAPDH).

    Techniques: Activity Assay

    TNF, IL-6 and MMP3 production (PB) in HC, SpA and ERA patients .  Scatter plots representing TNF, IL-6 and MMP3 production in PB in three group of subjects, HC (25), SpA (50) and ERA (52) patients as measured via ELISA. Each dot represents an individual sample. Horizontal line represents median. WB diluted 1:1 with complete culture medium was used for assessing the pro-inflammatory cytokine production post stimulation with endogenous and exogenous ligands. TNF production in response to ( a ). No stimulation ( b ). LPS or PG stimulation ( c ). TNC or MRP8 stimulation. IL-6 production in response to ( d ). No stimulation. ( e .) LPS or PG stimulation ( f ). TNC or MRP8 stimulation. MMP3 production in response to ( g ). No stimulation. ( h .) LPS or PG stimulation ( i ). TNC or MRP8 stimulation.  WB:  whole blood,  HC:  healthy controls,  SpA:  Spondyloarthropathy,  ERA:  enthesitis related arthritis,  Uns - unstimulated,  LPS-  Lipopolysaccaride,  PG-  peptidoglycan,  TNC-  Tenascin-C and  MRP8- Myeloid related protein 8,  TNF:  tumor necrosis factor,  IL-6:  Interleukin-6,  MMP3:  Matrix metalloproteinase 3

    Journal: Pediatric Rheumatology Online Journal

    Article Title: Patients with enthesitis related arthritis show similar monocyte function pattern as seen in adult axial spondyloarthropathy

    doi: 10.1186/s12969-020-0403-9

    Figure Lengend Snippet: TNF, IL-6 and MMP3 production (PB) in HC, SpA and ERA patients . Scatter plots representing TNF, IL-6 and MMP3 production in PB in three group of subjects, HC (25), SpA (50) and ERA (52) patients as measured via ELISA. Each dot represents an individual sample. Horizontal line represents median. WB diluted 1:1 with complete culture medium was used for assessing the pro-inflammatory cytokine production post stimulation with endogenous and exogenous ligands. TNF production in response to ( a ). No stimulation ( b ). LPS or PG stimulation ( c ). TNC or MRP8 stimulation. IL-6 production in response to ( d ). No stimulation. ( e .) LPS or PG stimulation ( f ). TNC or MRP8 stimulation. MMP3 production in response to ( g ). No stimulation. ( h .) LPS or PG stimulation ( i ). TNC or MRP8 stimulation. WB: whole blood, HC: healthy controls, SpA: Spondyloarthropathy, ERA: enthesitis related arthritis, Uns - unstimulated, LPS- Lipopolysaccaride, PG- peptidoglycan, TNC- Tenascin-C and MRP8- Myeloid related protein 8, TNF: tumor necrosis factor, IL-6: Interleukin-6, MMP3: Matrix metalloproteinase 3

    Article Snippet: Taqman gene expression assay kits were purchased from Applied Biosystems (CA, USA), the IDs being Hs00174128_m1 (TNF), Hs00174131_m1 (IL-6) and HS02786624_g1 (GAPDH).

    Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

    TNF and IL-6 producing monocyte frequency (PB) in HC, SpA and ERA patients. The figure shows scatter plots representing frequency of TNF and IL-6 producing monocytes in PB in HC (25), SpA (50) and ERA (52) patients as analysed via flow cytometry. Each dot represents an individual sample. Horizontal line represents median. WB diluted 1:1 with complete culture medium was used for assessing the pro-inflammatory cytokine producing monocyte frequency. Frequency of TNF producing monocytes in response to ( a ). No stimulation ( b ). LPS or PG stimulation ( c ). TNC or MRP8 stimulation. Frequency of IL-6 producing monocytes in response to ( d ). No stimulation. ( e .) LPS or PG stimulation ( f ). TNC or MRP8 stimulation.  WB:  whole blood,  HC:  healthy controls,  SpA:  Spondyloarthropathy,  ERA:  enthesitis related arthritis,  Uns - unstimulated,  LPS-  Lipopolysaccaride,  PG-  peptidoglycan,  TNC-  Tenascin-C and  MRP8- Myeloid related protein 8,  TNF:  tumor necrosis factor,  IL-6:  Interleukin-6

    Journal: Pediatric Rheumatology Online Journal

    Article Title: Patients with enthesitis related arthritis show similar monocyte function pattern as seen in adult axial spondyloarthropathy

    doi: 10.1186/s12969-020-0403-9

    Figure Lengend Snippet: TNF and IL-6 producing monocyte frequency (PB) in HC, SpA and ERA patients. The figure shows scatter plots representing frequency of TNF and IL-6 producing monocytes in PB in HC (25), SpA (50) and ERA (52) patients as analysed via flow cytometry. Each dot represents an individual sample. Horizontal line represents median. WB diluted 1:1 with complete culture medium was used for assessing the pro-inflammatory cytokine producing monocyte frequency. Frequency of TNF producing monocytes in response to ( a ). No stimulation ( b ). LPS or PG stimulation ( c ). TNC or MRP8 stimulation. Frequency of IL-6 producing monocytes in response to ( d ). No stimulation. ( e .) LPS or PG stimulation ( f ). TNC or MRP8 stimulation. WB: whole blood, HC: healthy controls, SpA: Spondyloarthropathy, ERA: enthesitis related arthritis, Uns - unstimulated, LPS- Lipopolysaccaride, PG- peptidoglycan, TNC- Tenascin-C and MRP8- Myeloid related protein 8, TNF: tumor necrosis factor, IL-6: Interleukin-6

    Article Snippet: Taqman gene expression assay kits were purchased from Applied Biosystems (CA, USA), the IDs being Hs00174128_m1 (TNF), Hs00174131_m1 (IL-6) and HS02786624_g1 (GAPDH).

    Techniques: Flow Cytometry, Cytometry, Western Blot

    TNF and IL-6 producing monocytes in SFMC in SpA vs ERA patients. Scatter plots representing frequency of TNF and IL-6 producing monocytes in SFMC in SpA (10) and ERA (10) patients in response to exogenous and endogenous TLR4 ligands analysed via flow cytometry. Each dot represents an individual sample. Horizontal line represents median. 10 6  SFMC/ml in complete culture medium was used.  a . Frequency of TNF producing monocytes ( b ). Frequency of IL-6 producing monocytes.  WB:  whole blood,  SpA:  Spondyloarthropathy,  ERA:  enthesitis related arthritis  Uns - unstimulated,  LPS-  Lipopolysaccaride,  PG-  peptidoglycan,  TNC-  Tenascin-C and  MRP8- Myeloid related protein 8,  TNF:  tumor necrosis factor,  IL-6:  Interleukin-6

    Journal: Pediatric Rheumatology Online Journal

    Article Title: Patients with enthesitis related arthritis show similar monocyte function pattern as seen in adult axial spondyloarthropathy

    doi: 10.1186/s12969-020-0403-9

    Figure Lengend Snippet: TNF and IL-6 producing monocytes in SFMC in SpA vs ERA patients. Scatter plots representing frequency of TNF and IL-6 producing monocytes in SFMC in SpA (10) and ERA (10) patients in response to exogenous and endogenous TLR4 ligands analysed via flow cytometry. Each dot represents an individual sample. Horizontal line represents median. 10 6 SFMC/ml in complete culture medium was used. a . Frequency of TNF producing monocytes ( b ). Frequency of IL-6 producing monocytes. WB: whole blood, SpA: Spondyloarthropathy, ERA: enthesitis related arthritis Uns - unstimulated, LPS- Lipopolysaccaride, PG- peptidoglycan, TNC- Tenascin-C and MRP8- Myeloid related protein 8, TNF: tumor necrosis factor, IL-6: Interleukin-6

    Article Snippet: Taqman gene expression assay kits were purchased from Applied Biosystems (CA, USA), the IDs being Hs00174128_m1 (TNF), Hs00174131_m1 (IL-6) and HS02786624_g1 (GAPDH).

    Techniques: Flow Cytometry, Cytometry, Western Blot

    IL-6 is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Quantitative RT-PCR, Expressing, In Situ Hybridization, Immunohistochemistry, Real-time Polymerase Chain Reaction

    IL-6/STAT3 signaling is activated in tracheal epithelium during repair. ( A ) Schematic of the SO 2 injury model. After exposure to SO 2 , luminal cells die. Basal cells spread, proliferate, and generate early progenitors. These progenitors differentiate into ciliated and secretory cells, and repair is complete in 2 wk. ( B ) Longitudinal midline sections stained with antibodies to p-STAT3 (red) and p63 (green), a marker for basal cells. ( C ) Expression of p-STAT3 (red) and FOXJ1 (green) during epithelial repair. Note the coexpression of p-STAT3 and FOXJ1 at 3 dpi. (Scale bars: B and C .)

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6/STAT3 signaling is activated in tracheal epithelium during repair. ( A ) Schematic of the SO 2 injury model. After exposure to SO 2 , luminal cells die. Basal cells spread, proliferate, and generate early progenitors. These progenitors differentiate into ciliated and secretory cells, and repair is complete in 2 wk. ( B ) Longitudinal midline sections stained with antibodies to p-STAT3 (red) and p63 (green), a marker for basal cells. ( C ) Expression of p-STAT3 (red) and FOXJ1 (green) during epithelial repair. Note the coexpression of p-STAT3 and FOXJ1 at 3 dpi. (Scale bars: B and C .)

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Staining, Marker, Expressing

    Effect of IL-6/STAT3 on tracheal epithelial repair in vivo. ( A ) Schematic of gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) model. Floxed alleles are deleted, and the YFP reporter is activated in basal cells with three doses of Tmx. One week later, mice are exposed to SO 2 and tracheas are harvested at 6 dpi. ( B ) Representative midline sections of tracheas (ventral) stained with YFP (lineage label, green) and a-tub (ciliated cells, red) in control ( K5-CreER T2 ; Rosa-YFP ) and gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) mice. A similar analysis was carried out using antibodies to K5 for basal cells and SCGB1A1 and SCGB3A2 for secretory cells, respectively. ( C ) Percentage of total lineage-labeled cells (YFP + ) throughout the trachea that are ciliated, secretory, or basal cells. Blue and red bars show K5-CreER T2 ; Rosa-YFP and K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP , respectively. ( D ) FOXJ1 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( E ) SCGB3A2 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( F ) In Il-6 null mice, there is a reduction of ciliated cells (FOXJ1 + ) and an increase of secretory cells (SCGB3A2 + ) after SO 2 injury (4 dpi). * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Effect of IL-6/STAT3 on tracheal epithelial repair in vivo. ( A ) Schematic of gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) model. Floxed alleles are deleted, and the YFP reporter is activated in basal cells with three doses of Tmx. One week later, mice are exposed to SO 2 and tracheas are harvested at 6 dpi. ( B ) Representative midline sections of tracheas (ventral) stained with YFP (lineage label, green) and a-tub (ciliated cells, red) in control ( K5-CreER T2 ; Rosa-YFP ) and gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) mice. A similar analysis was carried out using antibodies to K5 for basal cells and SCGB1A1 and SCGB3A2 for secretory cells, respectively. ( C ) Percentage of total lineage-labeled cells (YFP + ) throughout the trachea that are ciliated, secretory, or basal cells. Blue and red bars show K5-CreER T2 ; Rosa-YFP and K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP , respectively. ( D ) FOXJ1 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( E ) SCGB3A2 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( F ) In Il-6 null mice, there is a reduction of ciliated cells (FOXJ1 + ) and an increase of secretory cells (SCGB3A2 + ) after SO 2 injury (4 dpi). * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: In Vivo, Mouse Assay, Staining, Labeling

    Effect of IL-6 on regeneration of human epithelium in ALI culture. ( A ) Schematic of ALI culture of primary HBE cells. ( B ) Whole-mount staining of day 21 cultures for ciliated (α-tubulin, green) and secretory (SCGB3A1, red) cells. Nuclei are blue (DAPI). (Scale bar: 100 μm.) ( C ) Quantification of whole-mount staining, shown as a fold change over untreated culture. The α-tubulin + or SCGB3A1 + cells were counted in four randomly chosen areas (0.18 mm 2 ) per filter. Values are mean ± SD for cultures from three different donors. * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Effect of IL-6 on regeneration of human epithelium in ALI culture. ( A ) Schematic of ALI culture of primary HBE cells. ( B ) Whole-mount staining of day 21 cultures for ciliated (α-tubulin, green) and secretory (SCGB3A1, red) cells. Nuclei are blue (DAPI). (Scale bar: 100 μm.) ( C ) Quantification of whole-mount staining, shown as a fold change over untreated culture. The α-tubulin + or SCGB3A1 + cells were counted in four randomly chosen areas (0.18 mm 2 ) per filter. Values are mean ± SD for cultures from three different donors. * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Staining

    IL-6 enhances Foxj1-GFP expression in the mouse tracheosphere culture assay. ( A ) Schematic of the assay. NGFR + basal cells from Foxj1-GFP tracheas were cultured in 50% Matrigel in 96-well inserts. ( Right ) Section of a typical sphere with acetylated tubulin + (a-tub) ciliated (magenta) and Splunc + secretory cells (green). IHC, immunohistochemistry. The effect of IL-6 ( B ) and STAT3 inhibitor ( C ) on Foxj1 -GFP expression is shown. Differential interference contrast images ( Upper ) and fluorescent images ( Lower ) of the same spheres are shown. ( D ) Quantification by FACS at day 11 of the percentage of GFP + cells in dissociated spheres treated with IL-6 (0, 1, and 10 ng/mL). ( E ) Quantification at different times of GFP + cells in spheres cultured with or without IL-6 (1 ng/mL). ( F ) Representative sections of spheres at day 14 treated with IL-6 ( Left , 10 ng/mL) or S3I-201 ( Right , 200 μM, days 4–7). Both sections were stained with antibodies to a-tub + (magenta) and Splunc + (green). * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 enhances Foxj1-GFP expression in the mouse tracheosphere culture assay. ( A ) Schematic of the assay. NGFR + basal cells from Foxj1-GFP tracheas were cultured in 50% Matrigel in 96-well inserts. ( Right ) Section of a typical sphere with acetylated tubulin + (a-tub) ciliated (magenta) and Splunc + secretory cells (green). IHC, immunohistochemistry. The effect of IL-6 ( B ) and STAT3 inhibitor ( C ) on Foxj1 -GFP expression is shown. Differential interference contrast images ( Upper ) and fluorescent images ( Lower ) of the same spheres are shown. ( D ) Quantification by FACS at day 11 of the percentage of GFP + cells in dissociated spheres treated with IL-6 (0, 1, and 10 ng/mL). ( E ) Quantification at different times of GFP + cells in spheres cultured with or without IL-6 (1 ng/mL). ( F ) Representative sections of spheres at day 14 treated with IL-6 ( Left , 10 ng/mL) or S3I-201 ( Right , 200 μM, days 4–7). Both sections were stained with antibodies to a-tub + (magenta) and Splunc + (green). * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Expressing, Cell Culture, Immunohistochemistry, FACS, Staining

    IL-6 enhances expression of cilia-related genes and inhibits Notch1 expression in mouse ALI culture. ( A ) Schematic of ALI culture of mouse tracheal epithelial cells. At day 7, IL-6 (10 ng/mL) was added to culture medium in the lower chamber. Cells were harvested after 6, 12, and 24 h, and total RNA was extracted. ( B ) Quantitative RT-PCR shows that IL-6 treatment promotes the expression of the known target gene Socs3 and ciliogenesis-related genes, such as Multicilin ( Mcidas) and Foxj1 . IL-6 treatment also inhibits Notch1 and promotes expression of Cdc20b , the host gene for miR-449a/b. No significant changes were observed in the expression of Notch2 , Dll1 , or Jagged1 . ( C ) ChIP assay shows that p-STAT3 binding to promoter regions of Socs3, Foxj1 , Mcidas , and Notch1 is increased after IL-6 stimulation. * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 enhances expression of cilia-related genes and inhibits Notch1 expression in mouse ALI culture. ( A ) Schematic of ALI culture of mouse tracheal epithelial cells. At day 7, IL-6 (10 ng/mL) was added to culture medium in the lower chamber. Cells were harvested after 6, 12, and 24 h, and total RNA was extracted. ( B ) Quantitative RT-PCR shows that IL-6 treatment promotes the expression of the known target gene Socs3 and ciliogenesis-related genes, such as Multicilin ( Mcidas) and Foxj1 . IL-6 treatment also inhibits Notch1 and promotes expression of Cdc20b , the host gene for miR-449a/b. No significant changes were observed in the expression of Notch2 , Dll1 , or Jagged1 . ( C ) ChIP assay shows that p-STAT3 binding to promoter regions of Socs3, Foxj1 , Mcidas , and Notch1 is increased after IL-6 stimulation. * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Binding Assay

    Model for regulation of ciliogenesis in airway epithelium by STAT3. ( Upper ) After injury, STAT3 in both basal cells and progenitors is activated by IL-6 secreted from PDGFRα + stromal cells. Ciliogenesis is likely promoted both at the level of cell fate determination and at the level of differentiation/maturation of the progenitors of multiciliated cells. ( Lower ) Schematic model for how STAT3 may directly regulate ciliogenesis-related genes during repair of the tracheal epithelium.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Model for regulation of ciliogenesis in airway epithelium by STAT3. ( Upper ) After injury, STAT3 in both basal cells and progenitors is activated by IL-6 secreted from PDGFRα + stromal cells. Ciliogenesis is likely promoted both at the level of cell fate determination and at the level of differentiation/maturation of the progenitors of multiciliated cells. ( Lower ) Schematic model for how STAT3 may directly regulate ciliogenesis-related genes during repair of the tracheal epithelium.

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques:

    MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; IL-6, interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.

    Journal: PLoS ONE

    Article Title: Melanocortins Contribute to Sequential Differentiation and Enucleation of Human Erythroblasts via Melanocortin Receptors 1, 2 and 5

    doi: 10.1371/journal.pone.0123232

    Figure Lengend Snippet: MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; IL-6, interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.

    Article Snippet: In the second passage (for differentiation; D0–D3 in ), cell stocks were thawed and cultured at 2 × 105 cells/ml in HPGM supplemented with 3 U/ml human EPO (Kyowa Hakko Kirin), 25 ng/ml SCF, 10 ng/ml recombinant human IL-3 (PeproTech), and 10 ng/ml recombinant human IL-6 (R & D Systems) for 3 days ( ).

    Techniques: In Vitro, Staining, Reverse Transcription Polymerase Chain Reaction

    WT-CCN1, but not DM-CCN1, induces IL-6 in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: WT-CCN1, but not DM-CCN1, induces IL-6 in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Purification, Incubation, Blocking Assay

    WT-CCN1, but not DM-CCN1, induces IL-6 in vivo (A) IL-6 mRNA in the colon of wild type and Ccn1 dm/dm mice treated with 3.5% DSS for 5 days was measured by qRT-PCR, and normalized to healthy wild type colon with cyclophilin E as internal reference. Data shown as mean ± SD; n =3–4; n.s., not significant. ** P

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: WT-CCN1, but not DM-CCN1, induces IL-6 in vivo (A) IL-6 mRNA in the colon of wild type and Ccn1 dm/dm mice treated with 3.5% DSS for 5 days was measured by qRT-PCR, and normalized to healthy wild type colon with cyclophilin E as internal reference. Data shown as mean ± SD; n =3–4; n.s., not significant. ** P

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: In Vivo, Mouse Assay, Quantitative RT-PCR

    Delivery of IL-6 enhances survival and mucosal repair of Ccn1 dm/dm mice by restoring IEC proliferation (A) Ccn1 dm/dm mice ( n =20) were injected i.p. with rIL-6 (100 ng) or vehicle control daily for 5 consecutive days after each cycle of 5% DSS feeding and were monitored for survival. ** P

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: Delivery of IL-6 enhances survival and mucosal repair of Ccn1 dm/dm mice by restoring IEC proliferation (A) Ccn1 dm/dm mice ( n =20) were injected i.p. with rIL-6 (100 ng) or vehicle control daily for 5 consecutive days after each cycle of 5% DSS feeding and were monitored for survival. ** P

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: Mouse Assay, Injection

    Overexpression of PARK2 inhibits osteosarcoma cell tube formation in vitro. NC was used as the control group. The formation of capillary-like structures was obviously suppressed by PARK2. The co-culture with IL-6 (25 ng/ml) or stattic (10 μM) was also evaluated ( a ). The western blot analysis of the expression of the proteins p-Jak2, p-STAT3, STAT3, and VEGF post co-culture with IL-6 (25 ng/ml, 30 min) or stattic (10 μM, 2 h) ( b ). Magnification, ×100 ( a ). Scale bar, 200 μm ( a ). * P

    Journal: Cell Death & Disease

    Article Title: PARK2 inhibits osteosarcoma cell growth through the JAK2/STAT3/VEGF signaling pathway

    doi: 10.1038/s41419-018-0401-8

    Figure Lengend Snippet: Overexpression of PARK2 inhibits osteosarcoma cell tube formation in vitro. NC was used as the control group. The formation of capillary-like structures was obviously suppressed by PARK2. The co-culture with IL-6 (25 ng/ml) or stattic (10 μM) was also evaluated ( a ). The western blot analysis of the expression of the proteins p-Jak2, p-STAT3, STAT3, and VEGF post co-culture with IL-6 (25 ng/ml, 30 min) or stattic (10 μM, 2 h) ( b ). Magnification, ×100 ( a ). Scale bar, 200 μm ( a ). * P

    Article Snippet: The cells (2 × 104 cell/well) were re-suspended with medium, seeded in each well, and incubated with PBS, recombinant interleukin-6 (IL-6, 25 ng/ml; Peprotech, USA) or stattic (10 μM; Selleck, USA) at 37 °C in 5% CO2 .

    Techniques: Over Expression, In Vitro, Co-Culture Assay, Western Blot, Expressing

    Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c IL-6 and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P

    Journal: Journal of Nanobiotechnology

    Article Title: Orally administered gold nanoparticles protect against colitis by attenuating Toll-like receptor 4- and reactive oxygen/nitrogen species-mediated inflammatory responses but could induce gut dysbiosis in mice

    doi: 10.1186/s12951-018-0415-5

    Figure Lengend Snippet: Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c IL-6 and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P

    Article Snippet: The Pierce BCA protein assay kit, 2′,7′-DCF diacetate, the enhanced chemiluminescence (ECL) detection kit, and sandwich enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-6 and TNF-α, DMEM, Dulbecco’s phosphate-buffered saline (DPBS) and Hank’s balanced salt solution (HBSS) were obtained from ThermoFisher Scientific (CA, USA).

    Techniques: Activity Assay

    Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a 6-well plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.

    Journal: PLoS ONE

    Article Title: CTDSP1 inhibitor rabeprazole regulates DNA-PKcs dependent topoisomerase I degradation and irinotecan drug resistance in colorectal cancer

    doi: 10.1371/journal.pone.0228002

    Figure Lengend Snippet: Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a 6-well plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.

    Article Snippet: ImmunoblottingCells cultured in 6-well plates were scraped into an ice-cold RIPA buffer.

    Techniques: Microscopy, Immunofluorescence, Confocal Microscopy

    Silencing of CTDSP1 enhances topoI degradation and irinotecan resistance. A , Cells transfected with CTDSP1 or control siRNA were lyzed and the cells’ lysates were immunoblotted with anti-CTDSP1 and anti-β-actin antibodies. B , HCT116-siRNA CTDSP1 or control siRNA, treated with 2.5 μM SN-38 were harvested after 90 and 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. Cells’ lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. C , EGFP was integrated with topoI in HCT116 cells using CRISPR/Cas9 system and CTDSP1 was knocked down in this cell line by siRNA. Cells were treated with 2.5uM SN-38 for 60 min and the topoI-EGFP signal was imaged by Leica SP5 confocal microscope. D , HCT116-siRNA-CTDSP1 or control siRNA were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by detecting the luminescence. E , HCT116-siRNA-CTDSP1 or control siRNA cells were plated in a 6-well plate and treated with various concentrations of SN-38 for 24 h. Then, 50 cells per well were plated in a 6-well plate. After 14 days, when colonies were apparent (right panel), colonies were counted and the relative number of colonies was determined (left panel).

    Journal: PLoS ONE

    Article Title: CTDSP1 inhibitor rabeprazole regulates DNA-PKcs dependent topoisomerase I degradation and irinotecan drug resistance in colorectal cancer

    doi: 10.1371/journal.pone.0228002

    Figure Lengend Snippet: Silencing of CTDSP1 enhances topoI degradation and irinotecan resistance. A , Cells transfected with CTDSP1 or control siRNA were lyzed and the cells’ lysates were immunoblotted with anti-CTDSP1 and anti-β-actin antibodies. B , HCT116-siRNA CTDSP1 or control siRNA, treated with 2.5 μM SN-38 were harvested after 90 and 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. Cells’ lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. C , EGFP was integrated with topoI in HCT116 cells using CRISPR/Cas9 system and CTDSP1 was knocked down in this cell line by siRNA. Cells were treated with 2.5uM SN-38 for 60 min and the topoI-EGFP signal was imaged by Leica SP5 confocal microscope. D , HCT116-siRNA-CTDSP1 or control siRNA were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by detecting the luminescence. E , HCT116-siRNA-CTDSP1 or control siRNA cells were plated in a 6-well plate and treated with various concentrations of SN-38 for 24 h. Then, 50 cells per well were plated in a 6-well plate. After 14 days, when colonies were apparent (right panel), colonies were counted and the relative number of colonies was determined (left panel).

    Article Snippet: ImmunoblottingCells cultured in 6-well plates were scraped into an ice-cold RIPA buffer.

    Techniques: Transfection, CRISPR, Microscopy

    Comparison of TNF- α and IL-6 production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Bogijetong Decoction and Its Selected Formulation Are Involved in Alleviating Neuropathic Pain in a Rat Model of Chronic Constrictive Injury

    doi: 10.1155/2018/2050636

    Figure Lengend Snippet: Comparison of TNF- α and IL-6 production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p

    Article Snippet: Primary antibodies used in the present study were anti-BDNF (1:1000, mouse, monoclonal, Abcam, Cambridge, MA, USA), anti-TrkB (1:800, Rabbit, polyclonal, Abcam), anti-GAP-43 (1:800, Rabbit, polyclonal, Abcam), anti-phopsho-Erk1/2 (1:1000, Rabbit, polyclonal, Cell Signaling, Danvers, MA), anti-IL-6 (1:800, mouse, monoclonal, Abcam), anti-TNF- α (1:800, Rabbit, polyclonal, Abcam), and anti-Actin (1:20000, mouse, Sigma) antibodies, and secondary antibodies were goat anti-mouse and goat anti-rabbit horseradish peroxidase- (HRP-) conjugated antibodies (1:1000, Cell Signaling).

    Techniques: Western Blot

    Schematics of proposed mechanisms underlying BGJTD- or BeD-modulated pain responses. According to our data, signaling of TNF- α and TrkB appeared to increase after nerve ligation (a). TNF- α , which were decreased by BGJTD treatment, may cause weakening in retrograde signaling of TNFR into the cell body, leading to alleviating pain response (b). After BeD treatment, p-Erk1/2 and IL-6 signals, induced from Schwann cells and macrophages, may transmit the signals of IL-6R into the neuronal cell body (soma), inducing JAK/STAT activation and regulation of target gene expression including the repression of TrkB gene expression (c). Consequently, retrograde TrkB signaling would be attenuated and lead to alleviate pain response.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Bogijetong Decoction and Its Selected Formulation Are Involved in Alleviating Neuropathic Pain in a Rat Model of Chronic Constrictive Injury

    doi: 10.1155/2018/2050636

    Figure Lengend Snippet: Schematics of proposed mechanisms underlying BGJTD- or BeD-modulated pain responses. According to our data, signaling of TNF- α and TrkB appeared to increase after nerve ligation (a). TNF- α , which were decreased by BGJTD treatment, may cause weakening in retrograde signaling of TNFR into the cell body, leading to alleviating pain response (b). After BeD treatment, p-Erk1/2 and IL-6 signals, induced from Schwann cells and macrophages, may transmit the signals of IL-6R into the neuronal cell body (soma), inducing JAK/STAT activation and regulation of target gene expression including the repression of TrkB gene expression (c). Consequently, retrograde TrkB signaling would be attenuated and lead to alleviate pain response.

    Article Snippet: Primary antibodies used in the present study were anti-BDNF (1:1000, mouse, monoclonal, Abcam, Cambridge, MA, USA), anti-TrkB (1:800, Rabbit, polyclonal, Abcam), anti-GAP-43 (1:800, Rabbit, polyclonal, Abcam), anti-phopsho-Erk1/2 (1:1000, Rabbit, polyclonal, Cell Signaling, Danvers, MA), anti-IL-6 (1:800, mouse, monoclonal, Abcam), anti-TNF- α (1:800, Rabbit, polyclonal, Abcam), and anti-Actin (1:20000, mouse, Sigma) antibodies, and secondary antibodies were goat anti-mouse and goat anti-rabbit horseradish peroxidase- (HRP-) conjugated antibodies (1:1000, Cell Signaling).

    Techniques: Ligation, Activation Assay, Expressing

    STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human IL-6 for 30 min. Whole-cell lysates

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: STAT3 nuclear import is independent of tyrosine phosphorylation and mediated by importin-?3

    doi: 10.1073/pnas.0501643102

    Figure Lengend Snippet: STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human IL-6 for 30 min. Whole-cell lysates

    Article Snippet: Recombinant murine or human IL-6 (BioSource International, Camarillo, CA), IL-6 soluble receptor (R & D Systems) and human EGF (Sigma) were used at 10 ng/ml.

    Techniques: Translocation Assay, Transfection

    Anti-IL-6R reduces the expression of IFN-γ in activated Xiap −/− Treg cells. a , b Anti-IL-6R decreases IFN-γ expression in re-stimulated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated with anti-CD3/CD28 and IL-2, or with additional IL-12 as indicated, in the absence or presence of anti-IL-6R (50 μg ml −1 ) for 4 days. iTreg cells were re-stimulated with TPA/A23187 for 24 h and IFN-γ production was determined by ELISA ( a ), or reactivated with TPA/A23187 for 5 h and the expressions of Foxp3 and IFN-γ were determined by intracellular staining ( b ). c , d Anti-IL-6R inhibits IFN-γ expression in human Treg cells. Control and human XIAP-knockdown iTreg cells were stimulated as in ( a , b ), with additional IL-6 as indicated, and secretion ( c ) or intracellular expression ( d ) of IFN-γ was determined. e Inability of anti-TNF or anti-IL-1R to inhibit the production of IFN-γ in activated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated, as described in ( a ), in the presence or absence of anti-TNF or anti-IL-1R (50 μg ml −1 each) for 4 days. Treg cells were re-stimulated with TPA/A23187 for 24 h and the secreted IFN-γ was determined by ELISA. f Anti-IL-6R rescues the impaired suppressive activity of Xiap −/− tTreg cells in vivo. CD45.2 + WT or Xiap −/− tTreg cells were co-transferred with CD45.1 + CD4 + CD25 - effector T cells into male CD45.1 + RagI −/− mice. Anti-IL-6R antibody (500 μg per mouse) was intraperitoneally administrated at day 0, followed by weekly dosing of 500 μg. The body weights of mice were monitored at the indicated time-points. *** P

    Journal: Nature Communications

    Article Title: IL-6 receptor blockade corrects defects of XIAP-deficient regulatory T cells

    doi: 10.1038/s41467-018-02862-4

    Figure Lengend Snippet: Anti-IL-6R reduces the expression of IFN-γ in activated Xiap −/− Treg cells. a , b Anti-IL-6R decreases IFN-γ expression in re-stimulated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated with anti-CD3/CD28 and IL-2, or with additional IL-12 as indicated, in the absence or presence of anti-IL-6R (50 μg ml −1 ) for 4 days. iTreg cells were re-stimulated with TPA/A23187 for 24 h and IFN-γ production was determined by ELISA ( a ), or reactivated with TPA/A23187 for 5 h and the expressions of Foxp3 and IFN-γ were determined by intracellular staining ( b ). c , d Anti-IL-6R inhibits IFN-γ expression in human Treg cells. Control and human XIAP-knockdown iTreg cells were stimulated as in ( a , b ), with additional IL-6 as indicated, and secretion ( c ) or intracellular expression ( d ) of IFN-γ was determined. e Inability of anti-TNF or anti-IL-1R to inhibit the production of IFN-γ in activated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated, as described in ( a ), in the presence or absence of anti-TNF or anti-IL-1R (50 μg ml −1 each) for 4 days. Treg cells were re-stimulated with TPA/A23187 for 24 h and the secreted IFN-γ was determined by ELISA. f Anti-IL-6R rescues the impaired suppressive activity of Xiap −/− tTreg cells in vivo. CD45.2 + WT or Xiap −/− tTreg cells were co-transferred with CD45.1 + CD4 + CD25 - effector T cells into male CD45.1 + RagI −/− mice. Anti-IL-6R antibody (500 μg per mouse) was intraperitoneally administrated at day 0, followed by weekly dosing of 500 μg. The body weights of mice were monitored at the indicated time-points. *** P

    Article Snippet: Recombinant human IL-6 and human TGF-β were purchased from Peprotech (Rocky Hill, NJ).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, In Vivo, Mouse Assay

    IL-6 promotes NEAT1 transcription through STAT3 and H3K4me3. (A) NEAT1_2 promoter methylation in the QGY-7703 cells following IL-6 stimulation as indicated time. The black arrow shows the unmethylated bands. M, methylated; U, unmethylated. IL-6 6h, cells were treated with IL-6 for 6 h. IL-6 10h, cells were treated with IL-6 for 10 h. (B) H3K4me3, H3K9me3, H3K27me3 or H4K20me3 modifications at the NEAT1 promoter in the QGY-7703 cells with or without IL-6 stimulation were detected by ChIP assay. 0-1k: the sequence of 0 to 1000bp upstream transcription start site (TSS). 1k-2k: the sequence of 1000 to 2000bp upstream TSS (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: IL-6 promotes NEAT1 transcription through STAT3 and H3K4me3. (A) NEAT1_2 promoter methylation in the QGY-7703 cells following IL-6 stimulation as indicated time. The black arrow shows the unmethylated bands. M, methylated; U, unmethylated. IL-6 6h, cells were treated with IL-6 for 6 h. IL-6 10h, cells were treated with IL-6 for 10 h. (B) H3K4me3, H3K9me3, H3K27me3 or H4K20me3 modifications at the NEAT1 promoter in the QGY-7703 cells with or without IL-6 stimulation were detected by ChIP assay. 0-1k: the sequence of 0 to 1000bp upstream transcription start site (TSS). 1k-2k: the sequence of 1000 to 2000bp upstream TSS (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Methylation, Chromatin Immunoprecipitation, Sequencing

    Protein-protein interaction analysis. (A) QGY-7703 cells were incubated with IL-6 for 10 h. The cell lysates were collected and immunoprecipitated with anti-STAT3 antibody or IgG. The immuno-precipitates were subjected to western blotting analysis of STAT3 and PRDX5. (B) Wild-type or truncated STAT3 vectors were constructed as indicated. Association of Flag-tagged STAT3 with myc-tagged PRDX5 was determined using co-IP in the transfected HEK293T cells. (C) Wild-type or fragments of PRDX5 was constructed as indicated. Association of myc-tagged PRDX5 and Flag-tagged STAT3 was determined using co-IP in the transfected HEK293T cells.

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: Protein-protein interaction analysis. (A) QGY-7703 cells were incubated with IL-6 for 10 h. The cell lysates were collected and immunoprecipitated with anti-STAT3 antibody or IgG. The immuno-precipitates were subjected to western blotting analysis of STAT3 and PRDX5. (B) Wild-type or truncated STAT3 vectors were constructed as indicated. Association of Flag-tagged STAT3 with myc-tagged PRDX5 was determined using co-IP in the transfected HEK293T cells. (C) Wild-type or fragments of PRDX5 was constructed as indicated. Association of myc-tagged PRDX5 and Flag-tagged STAT3 was determined using co-IP in the transfected HEK293T cells.

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Incubation, Immunoprecipitation, Western Blot, Construct, Co-Immunoprecipitation Assay, Transfection

    Paraspeckle destruction inhibits IL-6-induced STAT3 phosphorylation. Phosphorylation of STAT3 and total STAT3 were analyzed by Western blotting in (A) QGY-7703 cells or (B) HepG2 cells transfected with si-NEAT1_2 or si-NONO with IL-6 stimulation as indicated time. (C) mRNA expression of Bcl-2, c-myc and Mmp-2 was measured in QGY-7703 cells transfected with si-NEAT1_2 or si-NONO with IL-6 stimulation for indicated time (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: Paraspeckle destruction inhibits IL-6-induced STAT3 phosphorylation. Phosphorylation of STAT3 and total STAT3 were analyzed by Western blotting in (A) QGY-7703 cells or (B) HepG2 cells transfected with si-NEAT1_2 or si-NONO with IL-6 stimulation as indicated time. (C) mRNA expression of Bcl-2, c-myc and Mmp-2 was measured in QGY-7703 cells transfected with si-NEAT1_2 or si-NONO with IL-6 stimulation for indicated time (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Western Blot, Transfection, Expressing

    Paraspeckle promotes IL-6-induced DNA damage through trapping GSTP1. (A) QGY-7703 cells were transfected with empty vector, myc-PRDX5 or myc-GSTP1 plasmids stimulated with or without IL-6 for 10 h, and nuclear extracts were used to detect the level of 8-OHDG using a 8-hydroxy 2 deoxyguanosine ELISA kit (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: Paraspeckle promotes IL-6-induced DNA damage through trapping GSTP1. (A) QGY-7703 cells were transfected with empty vector, myc-PRDX5 or myc-GSTP1 plasmids stimulated with or without IL-6 for 10 h, and nuclear extracts were used to detect the level of 8-OHDG using a 8-hydroxy 2 deoxyguanosine ELISA kit (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    IL-6 increases paraspeckle formation in HCC cells. NEAT1 and NEAT1_2 expressions were quantified by Q-PCR in QGY-7703 cells (A) or HepG2 cells (B) with or without IL-6 stimulation. Normalized with GAPDH mRNA levels, and compared with 0h group (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: IL-6 increases paraspeckle formation in HCC cells. NEAT1 and NEAT1_2 expressions were quantified by Q-PCR in QGY-7703 cells (A) or HepG2 cells (B) with or without IL-6 stimulation. Normalized with GAPDH mRNA levels, and compared with 0h group (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Polymerase Chain Reaction

    The effects of paraspeckle destruction on HCC cell invasion, proliferation and apoptosis under IL-6 stimulation. (A) The invasive ability of QGY-7703/HepG2 HCC cells was evaluated by in vitro invasion assay after transfection of NC, si-NEAT1_2 or si-NONO with or without IL-6 stimulation for 48 h (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: The effects of paraspeckle destruction on HCC cell invasion, proliferation and apoptosis under IL-6 stimulation. (A) The invasive ability of QGY-7703/HepG2 HCC cells was evaluated by in vitro invasion assay after transfection of NC, si-NEAT1_2 or si-NONO with or without IL-6 stimulation for 48 h (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: In Vitro, Invasion Assay, Transfection

    Paraspeckle promotes IL-6-induced STAT3 phosphorylation through trapping PRDX5 mRNA in nucleus. (A) QGY-7703 cells were transfected with empty vector plasmids or pCMV-myc-PRDX5 plasmids and then treated with IL-6 for the indicated time. Phosphorylation of STAT3 and total STAT3 were examined by Western blot. (B) Q-PCR detection of NEAT1_2 or PRDX5 mRNA retrieved by anti-NONO antibody or anti-IgG antibody with or without IL-6 stimulation for 10 h in RIP assay (* p

    Journal: Oncoimmunology

    Article Title: NEAT1 paraspeckle promotes human hepatocellular carcinoma progression by strengthening IL-6/STAT3 signaling

    doi: 10.1080/2162402X.2018.1503913

    Figure Lengend Snippet: Paraspeckle promotes IL-6-induced STAT3 phosphorylation through trapping PRDX5 mRNA in nucleus. (A) QGY-7703 cells were transfected with empty vector plasmids or pCMV-myc-PRDX5 plasmids and then treated with IL-6 for the indicated time. Phosphorylation of STAT3 and total STAT3 were examined by Western blot. (B) Q-PCR detection of NEAT1_2 or PRDX5 mRNA retrieved by anti-NONO antibody or anti-IgG antibody with or without IL-6 stimulation for 10 h in RIP assay (* p

    Article Snippet: Cells were treated with or without human IL-6 (50ng/ml, Peprotech, USA) for the indicated time, and then harvested.

    Techniques: Transfection, Plasmid Preparation, Western Blot, Polymerase Chain Reaction

    The LPD prevents renal injuries, the impairment of autophagy and the activation of mTORC1 in the kidneys of Wistar fatty rats. ( a ) Representative microphotographs of Masson’s trichrome (MT) staining of glomeruli (scale bar: 100 μm) and the tubulo-interstitial area (scale bar: 1 mm) after the 24-week dietary intervention. ( b ) Glomerular fibrotic score ( n = 3) and ( c ) tubulo-interstitial fibrotic score ( n = 3), assessed using MT staining after the 24-week dietary intervention. mRNA expression of Col3 ( d ), Kim-1 ( e ), Cd68 ( f ), Ccl12 ( g ), Tlr4 ( h ) and Il6 ( i ) in the renal cortex, adjusted to 18 s levels, after the 24-week dietary intervention ( n = 6 each). ( j ) Representative photographs of immunoblotting for p62, β-actin, p-S6RP and S6RP in the renal cortex after the 24-week dietary intervention ( n = 3). Quantitative ratios of p62 to β-actin ( k ) and p-S6RP to S6RP ( l ) ( n = 3). Col3: type 3 collagen, Kim-1: kidney injury molecule-1, Ccl2: C-C motif chemokine ligand 2, Tlr4: Toll-like receptor 4, Il6: interleukin-6, p-S6RP: phospho-S6 ribosomal protein, S6RP: S6 ribosomal protein. The data are shown as the mean ± SD. * p

    Journal: Nutrition & Metabolism

    Article Title: A low-protein diet exerts a beneficial effect on diabetic status and prevents diabetic nephropathy in Wistar fatty rats, an animal model of type 2 diabetes and obesity

    doi: 10.1186/s12986-018-0255-1

    Figure Lengend Snippet: The LPD prevents renal injuries, the impairment of autophagy and the activation of mTORC1 in the kidneys of Wistar fatty rats. ( a ) Representative microphotographs of Masson’s trichrome (MT) staining of glomeruli (scale bar: 100 μm) and the tubulo-interstitial area (scale bar: 1 mm) after the 24-week dietary intervention. ( b ) Glomerular fibrotic score ( n = 3) and ( c ) tubulo-interstitial fibrotic score ( n = 3), assessed using MT staining after the 24-week dietary intervention. mRNA expression of Col3 ( d ), Kim-1 ( e ), Cd68 ( f ), Ccl12 ( g ), Tlr4 ( h ) and Il6 ( i ) in the renal cortex, adjusted to 18 s levels, after the 24-week dietary intervention ( n = 6 each). ( j ) Representative photographs of immunoblotting for p62, β-actin, p-S6RP and S6RP in the renal cortex after the 24-week dietary intervention ( n = 3). Quantitative ratios of p62 to β-actin ( k ) and p-S6RP to S6RP ( l ) ( n = 3). Col3: type 3 collagen, Kim-1: kidney injury molecule-1, Ccl2: C-C motif chemokine ligand 2, Tlr4: Toll-like receptor 4, Il6: interleukin-6, p-S6RP: phospho-S6 ribosomal protein, S6RP: S6 ribosomal protein. The data are shown as the mean ± SD. * p

    Article Snippet: TaqMan probes for type 3 collagen (Col3) (Product ID: Rn01437681), Cd68 (Rn01495634), interleukin-6 (Il6) (Rn01410330), C-C motif chemokine ligand 2 (Ccl2) (Rn00580555), Toll-like receptor 4 (Tlr4) (Rn00569848), kidney injury molecule-1 (Kim-1) (RN00597703) and uncoupling protein1 (Ucp1) (Rn00562126) were purchased from Thermo Fisher Scientific (Waltham, MA, USA) [ ].

    Techniques: Activation Assay, Staining, Expressing

    IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of rmIL-6. The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p

    Journal: Annals of neurology

    Article Title: Opposite Effect Of Inflammation on SVZ vs. Hippocampal Precursors In Brain Injury

    doi: 10.1002/ana.22473

    Figure Lengend Snippet: IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of rmIL-6. The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p

    Article Snippet: Wells were washed, blocked and incubated with either dilutions of rmIL-6 or samples overnight at 4° C. The next day wells were washed and biotinylated anti-IL-6 antibody (R & D Systems) added for 2 hours at 37°C followed by washes and then incubation with AP-conjugated Streptavidin for 1 hour at 37°C.

    Techniques:

    Pancreatic cancer-educated macrophages induced the upregulation of CD59 in cancer cells via the IL-6R/STAT3 pathway. a Gene expression profiling in AsPC-1 cells cocultured with THP-1 macrophages compared with AsPC-1 cells. b , c GO and KEGG pathway analyses of differentially expressed genes. d IL6-R was extremely elevated in the cocultured group compared with that in the control group. e AsPC-1 and BxPC-3 cells were cultured with THP-1 macrophages and analyzed for the level of total STAT3 or phosphorylated STAT3 (p-STAT3). f , g IL-6 expression in macrophages cocultured with AsPC-1 and BxPC-3 cells was detected by qRT-PCR, western blot and ELISA. h The CD59 expression and phosphorylation of STAT3 in pancreatic cancer cells incubated with various concentrations of recombinant IL-6 (0, 0.01, 1, 10, and 100 nM) were detected by western blot

    Journal: Cell Death & Disease

    Article Title: Pancreatic cancer-educated macrophages protect cancer cells from complement-dependent cytotoxicity by up-regulation of CD59

    doi: 10.1038/s41419-019-2065-4

    Figure Lengend Snippet: Pancreatic cancer-educated macrophages induced the upregulation of CD59 in cancer cells via the IL-6R/STAT3 pathway. a Gene expression profiling in AsPC-1 cells cocultured with THP-1 macrophages compared with AsPC-1 cells. b , c GO and KEGG pathway analyses of differentially expressed genes. d IL6-R was extremely elevated in the cocultured group compared with that in the control group. e AsPC-1 and BxPC-3 cells were cultured with THP-1 macrophages and analyzed for the level of total STAT3 or phosphorylated STAT3 (p-STAT3). f , g IL-6 expression in macrophages cocultured with AsPC-1 and BxPC-3 cells was detected by qRT-PCR, western blot and ELISA. h The CD59 expression and phosphorylation of STAT3 in pancreatic cancer cells incubated with various concentrations of recombinant IL-6 (0, 0.01, 1, 10, and 100 nM) were detected by western blot

    Article Snippet: The primary antibodies used were as follows: anti-GAPDH (FL-335; Santa Cruz Biotechnology), anti-CD59 (HPA026494, Sigma-Aldrich), anti-CD163 (ab182422, Abcam), anti-arginase 1(Arg-1) (16001-1-AP, Proteintech), anti-IFN-γ (ab9657, Abcam), anti-iNOS (ab3523, Abcam), anti-IL-6 (ab6672, Abcam), anti-STAT3 (D1B2J, Cell Signaling Technology), and anti-phospho-Stat3 (Ser727, Cell Signaling Technology).

    Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Recombinant

    The effects on CD59 expression of IL6-antibody, si-IL-6, siSTAT3 and the STAT3 inhibitors. a IL-6 neutralization in the coculture system by IL-6 antibody inhibited CD59 expression in response to THP-1 coculture to a certain degree, but not significantly. b IL-6 siRNAs downregulated the IL-6 expression of THP-1. c After IL-6 knockdown of macrophages, CD59 upregulation was reversed in the coculture system. d siSTAT3 pancreatic cancer cells cocultured with THP-1 macrophages expressed lower amounts of CD59 than that of the siNC group. e STAT3 inhibitor, AG490 significantly decreased the expression of CD59 in THP-1-cocultured pancreatic cancer cells in a concentration-dependent manner. f The CD59 levels of the different representative groups were detected by FCM. g , h CDC and apoptosis assay between the representative groups in the conditioned media (fresh human serum, diluted 2:5)

    Journal: Cell Death & Disease

    Article Title: Pancreatic cancer-educated macrophages protect cancer cells from complement-dependent cytotoxicity by up-regulation of CD59

    doi: 10.1038/s41419-019-2065-4

    Figure Lengend Snippet: The effects on CD59 expression of IL6-antibody, si-IL-6, siSTAT3 and the STAT3 inhibitors. a IL-6 neutralization in the coculture system by IL-6 antibody inhibited CD59 expression in response to THP-1 coculture to a certain degree, but not significantly. b IL-6 siRNAs downregulated the IL-6 expression of THP-1. c After IL-6 knockdown of macrophages, CD59 upregulation was reversed in the coculture system. d siSTAT3 pancreatic cancer cells cocultured with THP-1 macrophages expressed lower amounts of CD59 than that of the siNC group. e STAT3 inhibitor, AG490 significantly decreased the expression of CD59 in THP-1-cocultured pancreatic cancer cells in a concentration-dependent manner. f The CD59 levels of the different representative groups were detected by FCM. g , h CDC and apoptosis assay between the representative groups in the conditioned media (fresh human serum, diluted 2:5)

    Article Snippet: The primary antibodies used were as follows: anti-GAPDH (FL-335; Santa Cruz Biotechnology), anti-CD59 (HPA026494, Sigma-Aldrich), anti-CD163 (ab182422, Abcam), anti-arginase 1(Arg-1) (16001-1-AP, Proteintech), anti-IFN-γ (ab9657, Abcam), anti-iNOS (ab3523, Abcam), anti-IL-6 (ab6672, Abcam), anti-STAT3 (D1B2J, Cell Signaling Technology), and anti-phospho-Stat3 (Ser727, Cell Signaling Technology).

    Techniques: Expressing, Neutralization, Concentration Assay, Apoptosis Assay

    IL-6 induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Coincorporation of LpxL1 and PagL Mutant Lipopolysaccharides into Liposomes with Neisseria meningitidis Opacity Protein: Influence on Endotoxic and Adjuvant Activity ▿

    doi: 10.1128/CVI.00423-09

    Figure Lengend Snippet: IL-6 induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).

    Article Snippet: Cells were incubated for 16 to 18 h at 37°C in a humid atmosphere with 5% CO2 , and the IL-6 concentrations in the supernatants were determined in an enzyme-linked immunosorbent assay (ELISA) against mouse IL-6 according to the manufacturer's instructions (BD Bioscience P, San Diego, CA).

    Techniques:

    MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P

    Journal: PLoS ONE

    Article Title: Mesenchymal stem cells ameliorate experimental arthritis via expression of interleukin-1 receptor antagonist

    doi: 10.1371/journal.pone.0193086

    Figure Lengend Snippet: MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P

    Article Snippet: The sorted CD4+ T cells were cultured in RPMI 1640 medium supplemented with 10% FBS (Gibco) and stimulated with 1 μg/ml plate-bound anti-mouse CD3 (BD Biosciences), 2 μg/ml anti-mouse CD28 (BD Biosciences), 2 μg/ml anti-mouse IL-4 (R & D Systems, MN, USA), 2 μg/ml anti-mouse interferon-γ (IFN-γ) (R & D Systems), 20 ng/ml recombinant IL-6 (R & D Systems), and 2 ng/ml recombinant transforming growth factor-β1 (TGF-β1) (R & D Systems) for 3 days.

    Techniques: Expressing, Flow Cytometry, Cytometry, Isolation

    Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Journal:

    Article Title: Attenuation of monocyte proinflammatory cytokine responses to Neisseria meningitidis in children by erythropoietin

    doi: 10.1111/j.1365-2249.2008.03760.x

    Figure Lengend Snippet: Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Article Snippet: Because non-haematopoietic effects of erythropoietin require higher concentrations [ ] we employed human recombinant erythropoietin beta (NeoRecormon, Firma Roche, Grenzen-Wylach, Germany), diluted in RPMI-1640 in final concentrations of 100 and 500 U/ml respectively, which was added to the cultures 1 h prior to stimulation with heat-inactivated meningococcal strains at a final concentration of 1·2 × 108 per ml for 5 h. As a positive control, lipopolysaccharide (LPS) obtained from Escherichia coli ( E. coli 0127: B8; no. L3129; Sigma, Deisenhofen, Germany) was added at a concentration of 30 ng/ml for 5 h. Cells were washed in Hanks's balanced salt solution (HBSS) and resuspended in a buffer consisting of HBSS, 0·1% saponin (Riedel de Haen, Hanover, Germany) and 0·01 M HEPES buffer (Seromed Biochrome, Berlin, Germany); 200 µl aliquots of cells were added to tubes containing 0·5 µg/10 µl of monoclonal antibodies (mAbs) against CD14, tumour necrosis factor (TNF)-α and interleukin (IL)-6 (Becton Dickinson, Heidelberg, Germany).

    Techniques:

    Determination of levels of TNF-α and IL-6 in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial ELISA kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).

    Journal: Frontiers in Microbiology

    Article Title: Pichia pastoris-Expressed Bivalent Virus-Like Particulate Vaccine Induces Domain III-Focused Bivalent Neutralizing Antibodies without Antibody-Dependent Enhancement in Vivo

    doi: 10.3389/fmicb.2017.02644

    Figure Lengend Snippet: Determination of levels of TNF-α and IL-6 in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial ELISA kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).

    Article Snippet: TNF-α and IL-6 in the clarified small intestinal extracts were determined using commercial cytokine ELISA kits (cat# KMC3011 for TNF-α; cat# KMC0061 for IL-6) procured from Invitrogen as per the manufacturer’s instructions.

    Techniques: Mouse Assay, In Vitro, Negative Control, Enzyme-linked Immunosorbent Assay, Purification, Recombinant, Standard Deviation

    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Journal: Molecular Medicine Reports

    Article Title: Syringaresinol-di-O-β-D-glucoside, a phenolic compound from Polygonatum sibiricum, exhibits an antidiabetic and antioxidative effect on a streptozotocin-induced mouse model of diabetes

    doi: 10.3892/mmr.2018.9580

    Figure Lengend Snippet: Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Article Snippet: Insulin (cat. no. ab100578) and interleukin-6 (IL-6; cat. no. ab100712) ELISA kits, and transforming growth factor-β1 (TGF-β1; cat. no. ab9758) and GAPDH (cat. no. ab8245) antibodies, were purchased from Abcam (Cambridge, MA, USA).

    Techniques: Mouse Assay, Injection, Standard Deviation

    Western blot analysis of protein expression of IL-6, p-STAT3, survivin, STAT3, and VEGF. Protein levels of IL-6, Survivin, p-STAT3, STAT3, and VEGF in normal gastric and tumor tissue were determined using western blotting. Beta-actin was a loading control. Relative protein expression of IL-6 (A), VEGF (B), surviving (C), p-STAT3 (D) was normalized to of the corresponding beta-actin level. Positive immunoreactive bands were quantified densitometrically and expressed as IL-6, Survivin, p-STAT3, STAT3, and VEGF in optical density units, respectively. * P

    Journal: PLoS ONE

    Article Title: Activation of STAT3 in Human Gastric Cancer Cells via Interleukin (IL)-6-Type Cytokine Signaling Correlates with Clinical Implications

    doi: 10.1371/journal.pone.0075788

    Figure Lengend Snippet: Western blot analysis of protein expression of IL-6, p-STAT3, survivin, STAT3, and VEGF. Protein levels of IL-6, Survivin, p-STAT3, STAT3, and VEGF in normal gastric and tumor tissue were determined using western blotting. Beta-actin was a loading control. Relative protein expression of IL-6 (A), VEGF (B), surviving (C), p-STAT3 (D) was normalized to of the corresponding beta-actin level. Positive immunoreactive bands were quantified densitometrically and expressed as IL-6, Survivin, p-STAT3, STAT3, and VEGF in optical density units, respectively. * P

    Article Snippet: Western blotting was performed using an anti-IL-6, anti-VEGF, anti-survivin, anti-STAT3 and phosphorylated STAT3 (Tyr705) antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) [ ].

    Techniques: Western Blot, Expressing

    Expression of STAT3 in human gastric cancer tissues. The expression and localization of STAT3, IL-6, VEGF, and survivin in gastric cancer cells were determined using immunohistochemical staining. There was weak or negative expression of STAT3 in adjacent normal mucosa. However, there was strong expression of phosphorylated STAT3 in gastric cancer tissues. The STAT3 staining was mainly localized in the nuclei of tumor epithelial cells, which was indicated by numerous yellowish granules. STAT3 overexpression was associated with with increased expression of IL-6, surviving, and VEGF as well as with increased vessel density (Original magnification of A1-A3 and B1-B3, ×400; A4 and B4, ×200)..

    Journal: PLoS ONE

    Article Title: Activation of STAT3 in Human Gastric Cancer Cells via Interleukin (IL)-6-Type Cytokine Signaling Correlates with Clinical Implications

    doi: 10.1371/journal.pone.0075788

    Figure Lengend Snippet: Expression of STAT3 in human gastric cancer tissues. The expression and localization of STAT3, IL-6, VEGF, and survivin in gastric cancer cells were determined using immunohistochemical staining. There was weak or negative expression of STAT3 in adjacent normal mucosa. However, there was strong expression of phosphorylated STAT3 in gastric cancer tissues. The STAT3 staining was mainly localized in the nuclei of tumor epithelial cells, which was indicated by numerous yellowish granules. STAT3 overexpression was associated with with increased expression of IL-6, surviving, and VEGF as well as with increased vessel density (Original magnification of A1-A3 and B1-B3, ×400; A4 and B4, ×200)..

    Article Snippet: Western blotting was performed using an anti-IL-6, anti-VEGF, anti-survivin, anti-STAT3 and phosphorylated STAT3 (Tyr705) antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) [ ].

    Techniques: Expressing, Immunohistochemistry, Staining, Over Expression

    Sorafenib regulates the expression of ID1/p16/IL6. a RT-PCR was conducted in HepG2 cells treated with different concentrations of sorafenib for 24 h. b Western blot experiments were performed in HepG2 cells treated with different concentrations of sorafenib for 24 h. c Cell supernatant was collected to perform the IL6 ELISA assay. d After incubated with 5 μM sorafenib for 24 h, cells were stained with SA-β-gal. * p

    Journal: Cell Death & Disease

    Article Title: ID1-induced p16/IL6 axis activation contributes to the resistant of hepatocellular carcinoma cells to sorafenib

    doi: 10.1038/s41419-018-0926-x

    Figure Lengend Snippet: Sorafenib regulates the expression of ID1/p16/IL6. a RT-PCR was conducted in HepG2 cells treated with different concentrations of sorafenib for 24 h. b Western blot experiments were performed in HepG2 cells treated with different concentrations of sorafenib for 24 h. c Cell supernatant was collected to perform the IL6 ELISA assay. d After incubated with 5 μM sorafenib for 24 h, cells were stained with SA-β-gal. * p

    Article Snippet: Neutralizing antibody of IL6 (Cat No: MAB206) was purchased from R & D Systems.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Staining

    Relationship between sorafenib efficacy and cell senescence. a The sensitivity of five HCC cell lines to sorafenib was measured by MTT assay. Cells were seeded in 96-well plates. After overnight incubation, cells were treated with different concentrations of sorafenib (5, 10, 20, 40 μM) for 24 h, then MTT assay was performed. DMSO was used as the control. The results were shown as inhibition rate, which indicates the percentage of cell growth inhibition caused by sorafenib treatment. b IC50 value was calculated based on the MTT results. The value represents the drug concentration of inducing 50% growth suppression compared to control cells. c HepG2 and Hep3B cells were incubated with sorafenib for 24 h, cell apoptosis was evaluated through flow cytometry (upper). The apoptosis ratio was calculated as the early apoptosis (lower right quadrant) plus the late apoptosis (upper right quadrant) percentage (lower). d HepG2 and Hep3B cells were incubated with β-gal staining solution. Senescent cells exhibited blue staining (left). Percentages of SA-β-gal positive cells were calculated and exhibited as a histogram (right). e Expression level of p16 in HepG2 and Hep3B cell lines was tested by western blot (left) and qRT-PCR (right). f IL6 expression in HepG2 and Hep3B cell lines was measured by qRT-PCR (left) and ELISA (right). For ELISA experiment, cells were seeded in 6-well plates and cultured for 24 h, then cell supernatants were collected. * p

    Journal: Cell Death & Disease

    Article Title: ID1-induced p16/IL6 axis activation contributes to the resistant of hepatocellular carcinoma cells to sorafenib

    doi: 10.1038/s41419-018-0926-x

    Figure Lengend Snippet: Relationship between sorafenib efficacy and cell senescence. a The sensitivity of five HCC cell lines to sorafenib was measured by MTT assay. Cells were seeded in 96-well plates. After overnight incubation, cells were treated with different concentrations of sorafenib (5, 10, 20, 40 μM) for 24 h, then MTT assay was performed. DMSO was used as the control. The results were shown as inhibition rate, which indicates the percentage of cell growth inhibition caused by sorafenib treatment. b IC50 value was calculated based on the MTT results. The value represents the drug concentration of inducing 50% growth suppression compared to control cells. c HepG2 and Hep3B cells were incubated with sorafenib for 24 h, cell apoptosis was evaluated through flow cytometry (upper). The apoptosis ratio was calculated as the early apoptosis (lower right quadrant) plus the late apoptosis (upper right quadrant) percentage (lower). d HepG2 and Hep3B cells were incubated with β-gal staining solution. Senescent cells exhibited blue staining (left). Percentages of SA-β-gal positive cells were calculated and exhibited as a histogram (right). e Expression level of p16 in HepG2 and Hep3B cell lines was tested by western blot (left) and qRT-PCR (right). f IL6 expression in HepG2 and Hep3B cell lines was measured by qRT-PCR (left) and ELISA (right). For ELISA experiment, cells were seeded in 6-well plates and cultured for 24 h, then cell supernatants were collected. * p

    Article Snippet: Neutralizing antibody of IL6 (Cat No: MAB206) was purchased from R & D Systems.

    Techniques: MTT Assay, Incubation, Inhibition, Concentration Assay, Flow Cytometry, Cytometry, Staining, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture

    Regulation of p16 and IL6 on the inhibitory effect of sorafenib on cell viability in HCC. a HepG2 and Hep3B cells were transfected with p16 overexpression vector and siRNA targeting p16, respectively. After 24 h, cells were incubated with different concentrations of sorafenib for 24 h. MTT assay was conducted to test cell viability. b After cultured with normal medium (Ctrl) or serum-free medium (Starvation) for 24 h, HepG2 cells were stained with β-gal solution. In Starvation group, senescent cells that exhibited blue staining were frequently observed under microscope. c p16 expression was detected by western blot. d IL6 concentration in cell supernatant was measured by ELISA. e MTT assay was used to determine the different effects of sorafenib on cell viability in HepG2 with routine culture (control group) or serum depletion (starvation group). f , g After cultured with normal medium (Ctrl) or serum-free medium (Starvation), cells were transfected with siRNA targeting p16 for 24 h ( f ) or pretreated with IL6 neutralizing antibody (5 ng/ml) for 2 h ( g ), and then incubated with 10 μM sorafenib for 24 h. MTT assay was conducted to test cell viability. NC: negative control. h In a transwell co-culture system, parent HepG2 cells were seeded in upper chamber, and the cells transfected with pCMV-p16 or empty vector were seeded in the bottom of wells. After 48 h-co-culture, the supernatant in bottom cells was collected to detect IL6 concentration. i The parent HepG2 cells in upper chamber were incubated with different concentrations of sorafenib for 24 h. Then cell viability was tested by MTT. * p

    Journal: Cell Death & Disease

    Article Title: ID1-induced p16/IL6 axis activation contributes to the resistant of hepatocellular carcinoma cells to sorafenib

    doi: 10.1038/s41419-018-0926-x

    Figure Lengend Snippet: Regulation of p16 and IL6 on the inhibitory effect of sorafenib on cell viability in HCC. a HepG2 and Hep3B cells were transfected with p16 overexpression vector and siRNA targeting p16, respectively. After 24 h, cells were incubated with different concentrations of sorafenib for 24 h. MTT assay was conducted to test cell viability. b After cultured with normal medium (Ctrl) or serum-free medium (Starvation) for 24 h, HepG2 cells were stained with β-gal solution. In Starvation group, senescent cells that exhibited blue staining were frequently observed under microscope. c p16 expression was detected by western blot. d IL6 concentration in cell supernatant was measured by ELISA. e MTT assay was used to determine the different effects of sorafenib on cell viability in HepG2 with routine culture (control group) or serum depletion (starvation group). f , g After cultured with normal medium (Ctrl) or serum-free medium (Starvation), cells were transfected with siRNA targeting p16 for 24 h ( f ) or pretreated with IL6 neutralizing antibody (5 ng/ml) for 2 h ( g ), and then incubated with 10 μM sorafenib for 24 h. MTT assay was conducted to test cell viability. NC: negative control. h In a transwell co-culture system, parent HepG2 cells were seeded in upper chamber, and the cells transfected with pCMV-p16 or empty vector were seeded in the bottom of wells. After 48 h-co-culture, the supernatant in bottom cells was collected to detect IL6 concentration. i The parent HepG2 cells in upper chamber were incubated with different concentrations of sorafenib for 24 h. Then cell viability was tested by MTT. * p

    Article Snippet: Neutralizing antibody of IL6 (Cat No: MAB206) was purchased from R & D Systems.

    Techniques: Transfection, Over Expression, Plasmid Preparation, Incubation, MTT Assay, Cell Culture, Staining, Microscopy, Expressing, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Serum Depletion, Negative Control, Co-Culture Assay

    ID1-mediated chemo-resistance. a After transfection with si-ID1 for 48 h, MTT assay was employed to detect the efficacy of sorafenib in HepG2 cells with ID1 knockdown. b After transfection with pcDNA-ID1 for 24 h, MTT assay was employed to detect the efficacy of sorafenib in Hep3B cells with ID1overexpression. c After transfection, HepG2 cells were pretreated with neutralizing antibody against IL6 (5 ng/ml) for 2 h prior to co-treatment with sorafenib (5 μM) for 24 h following by MTT assay. d After transfection, Hep3B cells were pretreated with neutralizing antibody against IL6 (5 ng/ml) for 2 h prior to co-treatment with sorafenib (20 μM) for 24 h following by MTT assay. * p

    Journal: Cell Death & Disease

    Article Title: ID1-induced p16/IL6 axis activation contributes to the resistant of hepatocellular carcinoma cells to sorafenib

    doi: 10.1038/s41419-018-0926-x

    Figure Lengend Snippet: ID1-mediated chemo-resistance. a After transfection with si-ID1 for 48 h, MTT assay was employed to detect the efficacy of sorafenib in HepG2 cells with ID1 knockdown. b After transfection with pcDNA-ID1 for 24 h, MTT assay was employed to detect the efficacy of sorafenib in Hep3B cells with ID1overexpression. c After transfection, HepG2 cells were pretreated with neutralizing antibody against IL6 (5 ng/ml) for 2 h prior to co-treatment with sorafenib (5 μM) for 24 h following by MTT assay. d After transfection, Hep3B cells were pretreated with neutralizing antibody against IL6 (5 ng/ml) for 2 h prior to co-treatment with sorafenib (20 μM) for 24 h following by MTT assay. * p

    Article Snippet: Neutralizing antibody of IL6 (Cat No: MAB206) was purchased from R & D Systems.

    Techniques: Transfection, MTT Assay

    Negative correlation between ID1 and p16/IL6 axis in HCC. a , b HepG2 cells transfected with siRNA against ID1 (Si-ID1) for 48 h. a RNA was isolated to detect ID1 and p16 to evaluate knockdown efficiency and its impact on p16 mRNA expression. b IL6 concentrations were quantified by ELISA (left). Percentages of SA-β-gal positive cells were calculated (right). c , d Hep3B cells transfected with pcDNA-ID1 for 24 h. c ID1 overexpression efficiency and its impact on p16 mRNA expression were determined by qRT-PCR. d IL6 concentrations were quantified by ELISA (left). Percentages of SA-β-gal positive cells were calculated (right). e A negative correlation between ID1 and p16 mRNA expression was observed in 24 HCC patient samples. f The levels of IL6, which were measured by ELISA, were higher in HCC patients with low concentrations of ID than those with high ones. * p

    Journal: Cell Death & Disease

    Article Title: ID1-induced p16/IL6 axis activation contributes to the resistant of hepatocellular carcinoma cells to sorafenib

    doi: 10.1038/s41419-018-0926-x

    Figure Lengend Snippet: Negative correlation between ID1 and p16/IL6 axis in HCC. a , b HepG2 cells transfected with siRNA against ID1 (Si-ID1) for 48 h. a RNA was isolated to detect ID1 and p16 to evaluate knockdown efficiency and its impact on p16 mRNA expression. b IL6 concentrations were quantified by ELISA (left). Percentages of SA-β-gal positive cells were calculated (right). c , d Hep3B cells transfected with pcDNA-ID1 for 24 h. c ID1 overexpression efficiency and its impact on p16 mRNA expression were determined by qRT-PCR. d IL6 concentrations were quantified by ELISA (left). Percentages of SA-β-gal positive cells were calculated (right). e A negative correlation between ID1 and p16 mRNA expression was observed in 24 HCC patient samples. f The levels of IL6, which were measured by ELISA, were higher in HCC patients with low concentrations of ID than those with high ones. * p

    Article Snippet: Neutralizing antibody of IL6 (Cat No: MAB206) was purchased from R & D Systems.

    Techniques: Transfection, Isolation, Expressing, Enzyme-linked Immunosorbent Assay, Over Expression, Quantitative RT-PCR

    Reduced inflammatory cytokine expression in retina from Lodamin-administered mice, and summary of Lodamin’s activity on T cells. (A) On day 21 after IRBP immunization, retinas were harvested and total RNA was extracted from naive mice (unimmunized control), EAU control group and Lodamin-administered group. The expression of IL-6, TNF, IFN-γ, and IL-17A were analyzed by realtime qPCR (mean ± SEM, n = 5–8, *** p

    Journal: PLoS ONE

    Article Title: Suppression of Autoimmune Retinal Inflammation by an Antiangiogenic Drug

    doi: 10.1371/journal.pone.0066219

    Figure Lengend Snippet: Reduced inflammatory cytokine expression in retina from Lodamin-administered mice, and summary of Lodamin’s activity on T cells. (A) On day 21 after IRBP immunization, retinas were harvested and total RNA was extracted from naive mice (unimmunized control), EAU control group and Lodamin-administered group. The expression of IL-6, TNF, IFN-γ, and IL-17A were analyzed by realtime qPCR (mean ± SEM, n = 5–8, *** p

    Article Snippet: Quantitative real-time PCR (qPCR) was performed using the MX3005P device (Stratagene, La Jolla, CA). cDNA was amplified in a 10 µg/ml final reaction mix containing TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and corresponding TaqMan Gene Expression Assays (Mm99999064_m1 (IL-6), Mm99999068_m1 (TNF), Mm00439619_m1 (IL-17A), Mm99999071_m1 (IFN-γ) and Mm00607939_s1 (β-actin, Applied Biosystems).

    Techniques: Expressing, Mouse Assay, Activity Assay, Real-time Polymerase Chain Reaction

    Lodamin inhibits Th1/Th17 cell differentiation by inhibiting cell proliferation and reducing the production of proinflammatory cytokines. (A) Purified CD4+CD25- T cells from spleen were activated by anti-CD3 antibody for 3 days under Th1 or Th17 polarization conditions in the presence or absence of Lodamin (1 nM), followed by stimulation with PMA and ionomycin in the presence of Brefeldin A; all plots are gated on CD4+ cells. Polarization conditions: Th0; anti-IFN-g Ab (10ug/ml) and anti-IL-4 Ab (10ug/ml); Th1; IL-12 (10 ng/ml) and anti-IL-4 Ab (10 µg/ml); Th17; IL-6 (20 ng/ml), TGFβ (2 ng/ml), anti-IFNγ Ab (10 µg/ml) and anti-IL-4 Ab (10 µg/ml) (B) Lodamin inhibits hallmark cytokine of Th1/Th17 under each polarization condition. IFN-γ and IL-17 protein level as measured by ELISA from activated cells supernatants (as describe in panel A). (mean ± SEM, n = 3 , *** p

    Journal: PLoS ONE

    Article Title: Suppression of Autoimmune Retinal Inflammation by an Antiangiogenic Drug

    doi: 10.1371/journal.pone.0066219

    Figure Lengend Snippet: Lodamin inhibits Th1/Th17 cell differentiation by inhibiting cell proliferation and reducing the production of proinflammatory cytokines. (A) Purified CD4+CD25- T cells from spleen were activated by anti-CD3 antibody for 3 days under Th1 or Th17 polarization conditions in the presence or absence of Lodamin (1 nM), followed by stimulation with PMA and ionomycin in the presence of Brefeldin A; all plots are gated on CD4+ cells. Polarization conditions: Th0; anti-IFN-g Ab (10ug/ml) and anti-IL-4 Ab (10ug/ml); Th1; IL-12 (10 ng/ml) and anti-IL-4 Ab (10 µg/ml); Th17; IL-6 (20 ng/ml), TGFβ (2 ng/ml), anti-IFNγ Ab (10 µg/ml) and anti-IL-4 Ab (10 µg/ml) (B) Lodamin inhibits hallmark cytokine of Th1/Th17 under each polarization condition. IFN-γ and IL-17 protein level as measured by ELISA from activated cells supernatants (as describe in panel A). (mean ± SEM, n = 3 , *** p

    Article Snippet: Quantitative real-time PCR (qPCR) was performed using the MX3005P device (Stratagene, La Jolla, CA). cDNA was amplified in a 10 µg/ml final reaction mix containing TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and corresponding TaqMan Gene Expression Assays (Mm99999064_m1 (IL-6), Mm99999068_m1 (TNF), Mm00439619_m1 (IL-17A), Mm99999071_m1 (IFN-γ) and Mm00607939_s1 (β-actin, Applied Biosystems).

    Techniques: Cell Differentiation, Purification, Enzyme-linked Immunosorbent Assay