gene exp il6 mm00446190 m1  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Thermo Fisher gene exp il6 mm00446190 m1
    Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) <t>IL-6</t> levels after CLP. *p
    Gene Exp Il6 Mm00446190 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 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 1 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 mm00446190 m1 - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "Obesity enhances sepsis induced liver inflammation and injury in mice"

    Article Title: Obesity enhances sepsis induced liver inflammation and injury in mice

    Journal: Obesity (Silver Spring, Md.)

    doi: 10.1002/oby.21504

    Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) IL-6 levels after CLP. *p
    Figure Legend Snippet: Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) IL-6 levels after CLP. *p

    Techniques Used: Mouse Assay

    The expression of (A) Adiponectin Receptor 2 (B) IL-6 (C) TNFα and (D) PPARγ in WAT were measured by RT-qPCR after CLP. *p
    Figure Legend Snippet: The expression of (A) Adiponectin Receptor 2 (B) IL-6 (C) TNFα and (D) PPARγ in WAT were measured by RT-qPCR after CLP. *p

    Techniques Used: Expressing, Quantitative RT-PCR

    2) Product Images from "IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages"

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages

    Journal: Current Medicinal Chemistry

    doi: 10.2174/1573406413666170922095011

    3-3- diindolylmethane (DIM) effect on MCP-1, IL-6 and TNF-α production in supernatants of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW 264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane, treated with 20 µM, 40 µM, and 60 µM DIM during 24 h followed by 100 nM insulin for 20 min. Determinations were carried out in triplicate using individual enzyme-linked immunosorbent assay and expressed as the mean value ± SD *p
    Figure Legend Snippet: 3-3- diindolylmethane (DIM) effect on MCP-1, IL-6 and TNF-α production in supernatants of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW 264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane, treated with 20 µM, 40 µM, and 60 µM DIM during 24 h followed by 100 nM insulin for 20 min. Determinations were carried out in triplicate using individual enzyme-linked immunosorbent assay and expressed as the mean value ± SD *p

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

    3-3-diindolylmethane (DIM) effect on MCP-1, IL-6, and TNF -α mRNA relative expression of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane treated with 20 µM, 40 µM, and 60 µM DIM during 24 h, followed by 100 nM insulin for 20 min. Determinations were performed by qPCR. Results were normalized based on the expression of actin-ß gene and analyzed by the comparative relative expression CT (2 -∆∆C T ) method [ 31 ] *p
    Figure Legend Snippet: 3-3-diindolylmethane (DIM) effect on MCP-1, IL-6, and TNF -α mRNA relative expression of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane treated with 20 µM, 40 µM, and 60 µM DIM during 24 h, followed by 100 nM insulin for 20 min. Determinations were performed by qPCR. Results were normalized based on the expression of actin-ß gene and analyzed by the comparative relative expression CT (2 -∆∆C T ) method [ 31 ] *p

    Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    3) Product Images from "Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury"

    Article Title: Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-020-01922-1

    Inflammatory cytokines. a Immunohistochemistry staining for inflammatory markers TNF-α and IL-6 in kidney tissue. b Western blot on kidney tissue measuring inflammatory markers TNF-α and NF-κB (left), alongside their quantification (right). c Quantitative real-time PCR on kidney tissue measuring inflammatory markers Nfkb , Il6 , and Tnfa . d ELISA measurement of blood serum concentrations of cytokines IL-1β, IL-6, and TNF-α. Each group has n = 5 mice. Significant difference a p
    Figure Legend Snippet: Inflammatory cytokines. a Immunohistochemistry staining for inflammatory markers TNF-α and IL-6 in kidney tissue. b Western blot on kidney tissue measuring inflammatory markers TNF-α and NF-κB (left), alongside their quantification (right). c Quantitative real-time PCR on kidney tissue measuring inflammatory markers Nfkb , Il6 , and Tnfa . d ELISA measurement of blood serum concentrations of cytokines IL-1β, IL-6, and TNF-α. Each group has n = 5 mice. Significant difference a p

    Techniques Used: Immunohistochemistry, Staining, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Mouse Assay

    4) Product Images from "IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content"

    Article Title: IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0051233

    Mitochondrial content and respiration are not altered in epididymal adipose tissue from IL-6 −/− mice fed a HFD. A) Mitochondrial marker proteins, B) mtDNA content and C) mitochondrial respiration are similar between genotypes. Data are presented as means + SE for 8–10 animals per group. Western blot images are given above the quantified data in A. Relative mtDNA content is normalized to genomic MCT1 (monocarboxylate transporter 1) DNA content and expressed as fold differences compared to the WT group in B. Mitochondrial respiration was measured by high-resolution O2 consumption. GM (glutamate, malate), GMD (glutamate, malate, ADP), GMDPS (glutamate, malate, ADP, pyruvate, succinate).
    Figure Legend Snippet: Mitochondrial content and respiration are not altered in epididymal adipose tissue from IL-6 −/− mice fed a HFD. A) Mitochondrial marker proteins, B) mtDNA content and C) mitochondrial respiration are similar between genotypes. Data are presented as means + SE for 8–10 animals per group. Western blot images are given above the quantified data in A. Relative mtDNA content is normalized to genomic MCT1 (monocarboxylate transporter 1) DNA content and expressed as fold differences compared to the WT group in B. Mitochondrial respiration was measured by high-resolution O2 consumption. GM (glutamate, malate), GMD (glutamate, malate, ADP), GMDPS (glutamate, malate, ADP, pyruvate, succinate).

    Techniques Used: Mouse Assay, Marker, Western Blot

    IL-6 exerts depot specific differences in gene expression in mouse adipose tissue. IL-6 treatment (6 hours, 75 ng/ml) increased the expression of A) SOCS3, PGC-1α and PRC mRNA in cultured epididymal adipose tissue (eWAT). B) A 12 hr IL-6 (75 ng/ml) treatment increased the expression of COXIV and CPT-1 in cultured eWAT. C) Cultured subcutaneous adipose tissue (sWAT) was not responsive to IL-6 treatment (75 ng/ml, 6 hours) and this was associated with D) reductions in the protein content of IL-6 receptor alpha and GP130. Data are presented as means + SE for 7 cultures, each from an individual mouse, per group and is expressed relative to the vehicle treated control culture from the same animal. For the Western blot data in D) data are presented as means + SE for 8–10 mice per group. Representative Western blots are presented above the quantified data. * P
    Figure Legend Snippet: IL-6 exerts depot specific differences in gene expression in mouse adipose tissue. IL-6 treatment (6 hours, 75 ng/ml) increased the expression of A) SOCS3, PGC-1α and PRC mRNA in cultured epididymal adipose tissue (eWAT). B) A 12 hr IL-6 (75 ng/ml) treatment increased the expression of COXIV and CPT-1 in cultured eWAT. C) Cultured subcutaneous adipose tissue (sWAT) was not responsive to IL-6 treatment (75 ng/ml, 6 hours) and this was associated with D) reductions in the protein content of IL-6 receptor alpha and GP130. Data are presented as means + SE for 7 cultures, each from an individual mouse, per group and is expressed relative to the vehicle treated control culture from the same animal. For the Western blot data in D) data are presented as means + SE for 8–10 mice per group. Representative Western blots are presented above the quantified data. * P

    Techniques Used: Expressing, Pyrolysis Gas Chromatography, Cell Culture, Cycling Probe Technology, Western Blot, Mouse Assay

    PGC-1 expression and mitochondrial markers proteins are not different in adipose tissue from WT and IL-6 −/− mice. The mRNA expression of PGC-1 co-activators (A and B) and mitochondrial marker proteins (C and D) are not different in epididymal (A and C) and subcutaneous adipose tissue (B and D) from WT and IL-6 −/− deficient mice. In A and B mRNA data is expressed relative to values in WT controls. Representative Western blots are given above the quantified data in C and D. Data are means + SE for 10 samples per group.
    Figure Legend Snippet: PGC-1 expression and mitochondrial markers proteins are not different in adipose tissue from WT and IL-6 −/− mice. The mRNA expression of PGC-1 co-activators (A and B) and mitochondrial marker proteins (C and D) are not different in epididymal (A and C) and subcutaneous adipose tissue (B and D) from WT and IL-6 −/− deficient mice. In A and B mRNA data is expressed relative to values in WT controls. Representative Western blots are given above the quantified data in C and D. Data are means + SE for 10 samples per group.

    Techniques Used: Pyrolysis Gas Chromatography, Expressing, Mouse Assay, Marker, Western Blot

    Glucose and insulin tolerance in WT and IL-6 −/− mice fed a HFD. Glucose curves and the quantified area under the curves for glucose (A, B) and insulin (C, D) tolerance tests. Data are presented as means +SE for 8–10 mice per group. * P
    Figure Legend Snippet: Glucose and insulin tolerance in WT and IL-6 −/− mice fed a HFD. Glucose curves and the quantified area under the curves for glucose (A, B) and insulin (C, D) tolerance tests. Data are presented as means +SE for 8–10 mice per group. * P

    Techniques Used: Mouse Assay

    5) Product Images from "Cardiac Function Remains Impaired Despite Reversible Cardiac Remodeling after Acute Experimental Viral Myocarditis"

    Article Title: Cardiac Function Remains Impaired Despite Reversible Cardiac Remodeling after Acute Experimental Viral Myocarditis

    Journal: Journal of Immunology Research

    doi: 10.1155/2017/6590609

    Cytokine expression in cardiac tissue and serum of CVB3-infected C57BL/6J mice 7 and 28 days after infection. (a) Cardiac tissue of CVB3-infected C57BL/6J mice was used for TaqMan based gene expression analysis of various chemotactic chemokines and cytokines. The gene expression of chemokines (Ccl2 , Ccl5 , Ccl7 , Cxcl10, and Cxcl13) as well as the expression of Il-6 and Tnf-α was highly increased 7 days p.i. compared to healthy controls and returned to basal levels 28 days after infection. Gene expression of Il-23a expression was not altered in cardiac tissue after CVB3 infection. Data are presented as absolute mRNA expression ( x -fold to the house keeping gene 18S) in box plots as well as in fold change to control animals as mean ± SEM above the corresponding bar using the formula 2 −ΔΔCt . (b) Protein expression in cardiac tissue was determined using Bioplex. The protein expression of MCP-1 and RANTES was increased 7 days after infection and dropped down 28 days after infection. (c) Protein expression in serum was detected using protein profiler arrays. The chemokines MCP-1, RANTES, IP-10, and CXCL13 were increased in serum samples of CVB3 infected mice 7 days after infection. Expression levels from noninfected control mice are shown as white bars, from CVB3-infected animals 7 days p.i. as light grey bars and from CVB3-infected animals 28 days p.i. as dark-grey bars; ∗ significantly different compared to noninfected mice (control); ∗ P
    Figure Legend Snippet: Cytokine expression in cardiac tissue and serum of CVB3-infected C57BL/6J mice 7 and 28 days after infection. (a) Cardiac tissue of CVB3-infected C57BL/6J mice was used for TaqMan based gene expression analysis of various chemotactic chemokines and cytokines. The gene expression of chemokines (Ccl2 , Ccl5 , Ccl7 , Cxcl10, and Cxcl13) as well as the expression of Il-6 and Tnf-α was highly increased 7 days p.i. compared to healthy controls and returned to basal levels 28 days after infection. Gene expression of Il-23a expression was not altered in cardiac tissue after CVB3 infection. Data are presented as absolute mRNA expression ( x -fold to the house keeping gene 18S) in box plots as well as in fold change to control animals as mean ± SEM above the corresponding bar using the formula 2 −ΔΔCt . (b) Protein expression in cardiac tissue was determined using Bioplex. The protein expression of MCP-1 and RANTES was increased 7 days after infection and dropped down 28 days after infection. (c) Protein expression in serum was detected using protein profiler arrays. The chemokines MCP-1, RANTES, IP-10, and CXCL13 were increased in serum samples of CVB3 infected mice 7 days after infection. Expression levels from noninfected control mice are shown as white bars, from CVB3-infected animals 7 days p.i. as light grey bars and from CVB3-infected animals 28 days p.i. as dark-grey bars; ∗ significantly different compared to noninfected mice (control); ∗ P

    Techniques Used: Expressing, Infection, Mouse Assay

    6) Product Images from "Acute and chronic effects of exercise on mRNA expression in the skeletal muscle of two mouse models of peripheral artery disease"

    Article Title: Acute and chronic effects of exercise on mRNA expression in the skeletal muscle of two mouse models of peripheral artery disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0182456

    Acute effects of treadmill exercise on the mRNA expression of known exercise-responsive genes. The soleus muscles of exercised C57BL/6 PAD mice (n = 4 per time point, total 24, left panels) and KK- A y PAD mice (n = 6 per time point, total 30, right panels) were obtained before and after the fifth treadmill exercise session. The mRNA expression levels of Pgc1a (A), Il6 (B), Nr4a1 (C), Nr4a2 (D), and Nr4a3 (E) in the soleus muscles were determined by qRT-PCR, normalized to Rplp0 expression level, and calculated relative to the pre-exercise group (0 min). The results are expressed as the mean ± SD. * p
    Figure Legend Snippet: Acute effects of treadmill exercise on the mRNA expression of known exercise-responsive genes. The soleus muscles of exercised C57BL/6 PAD mice (n = 4 per time point, total 24, left panels) and KK- A y PAD mice (n = 6 per time point, total 30, right panels) were obtained before and after the fifth treadmill exercise session. The mRNA expression levels of Pgc1a (A), Il6 (B), Nr4a1 (C), Nr4a2 (D), and Nr4a3 (E) in the soleus muscles were determined by qRT-PCR, normalized to Rplp0 expression level, and calculated relative to the pre-exercise group (0 min). The results are expressed as the mean ± SD. * p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR

    7) Product Images from "Attenuated viral hepatitis in Trem1−/− mice is associated with reduced inflammatory activity of neutrophils"

    Article Title: Attenuated viral hepatitis in Trem1−/− mice is associated with reduced inflammatory activity of neutrophils

    Journal: Scientific Reports

    doi: 10.1038/srep28556

    Impaired secretion of CCL2 and TNF-α by TREM1−/− neutrophils in response to LCMV. ( A ) Neutrophils were isolated from Trem1 +/+ C57BL/6 mice, stimulated with LCMV WE (MOI 5) and, after 6 hours, the expression of CCL2, TNF-α, IL-1β, IL-6, MPO, CXCL1, CXCL2, CXCL5, IFN-α and IFN-γ relative to the HPRT house-keeper were determined by qRT-PCR. Shown are mean values ± SEM. *p
    Figure Legend Snippet: Impaired secretion of CCL2 and TNF-α by TREM1−/− neutrophils in response to LCMV. ( A ) Neutrophils were isolated from Trem1 +/+ C57BL/6 mice, stimulated with LCMV WE (MOI 5) and, after 6 hours, the expression of CCL2, TNF-α, IL-1β, IL-6, MPO, CXCL1, CXCL2, CXCL5, IFN-α and IFN-γ relative to the HPRT house-keeper were determined by qRT-PCR. Shown are mean values ± SEM. *p

    Techniques Used: Isolation, Mouse Assay, Expressing, Quantitative RT-PCR

    8) Product Images from "Isothiocyanate-enriched moringa seed extract alleviates ulcerative colitis symptoms in mice"

    Article Title: Isothiocyanate-enriched moringa seed extract alleviates ulcerative colitis symptoms in mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0184709

    Effects of MSE on colonic expression of pro-inflammatory markers, tight-junction proteins, and Nrf2-mediated enzymes in DSS-induced UC. Gene expression of (A) IL-1β, (B) IL-6, (C) TNF-α, (D) iNOS, (E) claudin-1, (F) ZO-1, (G) GSTP1, (H) NQO1, and (I) HO1 was analyzed by qPCR in colon tissues of acute UC (n = 8/group) and chronic UC (n = 6/group) mice. All data are expressed as a relative fold change compared to healthy control (value = 1.0). Experimental designs and disease induction procedures are defined in Table 1 . Data are expressed as mean ± SEM. Different letters (a, b) indicate significant differences between groups at p
    Figure Legend Snippet: Effects of MSE on colonic expression of pro-inflammatory markers, tight-junction proteins, and Nrf2-mediated enzymes in DSS-induced UC. Gene expression of (A) IL-1β, (B) IL-6, (C) TNF-α, (D) iNOS, (E) claudin-1, (F) ZO-1, (G) GSTP1, (H) NQO1, and (I) HO1 was analyzed by qPCR in colon tissues of acute UC (n = 8/group) and chronic UC (n = 6/group) mice. All data are expressed as a relative fold change compared to healthy control (value = 1.0). Experimental designs and disease induction procedures are defined in Table 1 . Data are expressed as mean ± SEM. Different letters (a, b) indicate significant differences between groups at p

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

    9) Product Images from "Kallistatin attenuates endothelial senescence by modulating Let‐7g‐mediated miR‐34a‐ SIRT1‐ eNOS pathway. Kallistatin attenuates endothelial senescence by modulating Let‐7g‐mediated miR‐34a‐SIRT1‐eNOS pathway"

    Article Title: Kallistatin attenuates endothelial senescence by modulating Let‐7g‐mediated miR‐34a‐ SIRT1‐ eNOS pathway. Kallistatin attenuates endothelial senescence by modulating Let‐7g‐mediated miR‐34a‐SIRT1‐eNOS pathway

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13734

    Kallistatin ( KS ) deficiency in mouse lung endothelial cells increases senescence, oxidative stress and inflammation. Endothelial marker  CD 31 expression in mouse endothelial cells determined by  PCR  (A) (n = 3/group). Representative images of immunostaining of  CD 31 (green), with nuclei (Hoechst 33342, blue) and their merged images are shown (B) (n = 3/group). Identification of mouse kallistatin depletion (del) by target null gene  DNA  electrophoresis (C) (n = 3/group). Representative immunoblots of mouse  KS  protein (D) (n = 3/group), and mouse  KS mRNA  levels (E) (n = 6/group). Quantitative analysis of  SA ‐β‐gal activity (F), p16 INK 4a  and  PAI ‐1  mRNA  levels (G, H), superoxide formation (I),  NADPH  oxidase activity (J), VCAM ‐1,  ICAM ‐1 and  IL ‐6  mRNA  levels (K‐M) in mouse endothelial cells with or without H 2 O 2  treatment (n = 6/group). Values are expressed as mean ± SEM. * P
    Figure Legend Snippet: Kallistatin ( KS ) deficiency in mouse lung endothelial cells increases senescence, oxidative stress and inflammation. Endothelial marker CD 31 expression in mouse endothelial cells determined by PCR (A) (n = 3/group). Representative images of immunostaining of CD 31 (green), with nuclei (Hoechst 33342, blue) and their merged images are shown (B) (n = 3/group). Identification of mouse kallistatin depletion (del) by target null gene DNA electrophoresis (C) (n = 3/group). Representative immunoblots of mouse KS protein (D) (n = 3/group), and mouse KS mRNA levels (E) (n = 6/group). Quantitative analysis of SA ‐β‐gal activity (F), p16 INK 4a and PAI ‐1 mRNA levels (G, H), superoxide formation (I), NADPH oxidase activity (J), VCAM ‐1, ICAM ‐1 and IL ‐6 mRNA levels (K‐M) in mouse endothelial cells with or without H 2 O 2 treatment (n = 6/group). Values are expressed as mean ± SEM. * P

    Techniques Used: Marker, Expressing, Polymerase Chain Reaction, Immunostaining, Nucleic Acid Electrophoresis, Western Blot, Activity Assay

    Signalling pathway by which kallistatin inhibits endothelial senescence, oxidative stress and inflammation. Kallistatin through tyrosine kinase stimulates Let‐7g synthesis, inhibits miR‐34 and up‐regulates  SIRT 1‐mediated‐ eNOS  pathway, thereby elevating catalase,  SOD ‐1/2 and  NO  levels, leading to blockade of H 2 O 2 ‐modulated  ROS  formation, p16 INK 4a ,  PAI ‐1,  VCAM ‐1,  ICAM ‐1 and  IL ‐6 expression, and telomerase activity in endothelial cells. Genistein, tyrosine kinase inhibitor;  NAM ,  SIRT 1 inhibitor; L‐ NAME ,  NOS  inhibitor
    Figure Legend Snippet: Signalling pathway by which kallistatin inhibits endothelial senescence, oxidative stress and inflammation. Kallistatin through tyrosine kinase stimulates Let‐7g synthesis, inhibits miR‐34 and up‐regulates SIRT 1‐mediated‐ eNOS pathway, thereby elevating catalase, SOD ‐1/2 and NO levels, leading to blockade of H 2 O 2 ‐modulated ROS formation, p16 INK 4a , PAI ‐1, VCAM ‐1, ICAM ‐1 and IL ‐6 expression, and telomerase activity in endothelial cells. Genistein, tyrosine kinase inhibitor; NAM , SIRT 1 inhibitor; L‐ NAME , NOS inhibitor

    Techniques Used: Expressing, Activity Assay

    Kallistatin ( KS ) suppresses H 2 O 2 ‐induced oxidative stress and inflammation in human endothelial cells. Representative images of superoxide formation determined by fluorescence dye  DCF  (A), and quantitative analysis of  DCF  fluorescence (B).  NADPH  oxidase activity (C).  mRNA  levels of  NADPH  oxidase 4 (D),  VCAM ‐1 (E),  ICAM ‐1 (F),  IL ‐6 (G) and miR‐34a (H) analysed by  qRT ‐ PCR . Values are expressed as mean ± SEM. (n = 3 in each group). * P
    Figure Legend Snippet: Kallistatin ( KS ) suppresses H 2 O 2 ‐induced oxidative stress and inflammation in human endothelial cells. Representative images of superoxide formation determined by fluorescence dye DCF (A), and quantitative analysis of DCF fluorescence (B). NADPH oxidase activity (C). mRNA levels of NADPH oxidase 4 (D), VCAM ‐1 (E), ICAM ‐1 (F), IL ‐6 (G) and miR‐34a (H) analysed by qRT ‐ PCR . Values are expressed as mean ± SEM. (n = 3 in each group). * P

    Techniques Used: Fluorescence, Activity Assay, Quantitative RT-PCR

    10) Product Images from "Obesity exacerbates colitis-associated cancer via IL-6-regulated macrophage polarisation and CCL-20/CCR-6-mediated lymphocyte recruitment"

    Article Title: Obesity exacerbates colitis-associated cancer via IL-6-regulated macrophage polarisation and CCL-20/CCR-6-mediated lymphocyte recruitment

    Journal: Nature Communications

    doi: 10.1038/s41467-018-03773-0

    Mechanism how obesity and IL-6 alter the colitis-associated tumour microenvironment. 1. Obesity-induced gut barrier defects cause a low-grade inflammation including increases in IL-6 levels. 2. IL-6 polarises macrophages towards M2-type in colitis-associated colorectal cancer (CAC) that in response to commensal factors express the chemoattractant CCL-20. 3. CCL-20 recruits CCR-6-expressing lymphocytes comprising B cells, regulatory T cells (Treg) and T cells expressing αβ and γδ T-cell receptors that impact on CAC progression. B cells impact on macrophage polarisation, whereas CCR-6 + γδ T cells provide pathogenic IL-17. 4. IL-6 signalling in αβ T cells releases them from Treg-mediated suppression to exert their effector functions such as T helper (Th)1 and Th17 responses. TAM, tumour-associated macrophage
    Figure Legend Snippet: Mechanism how obesity and IL-6 alter the colitis-associated tumour microenvironment. 1. Obesity-induced gut barrier defects cause a low-grade inflammation including increases in IL-6 levels. 2. IL-6 polarises macrophages towards M2-type in colitis-associated colorectal cancer (CAC) that in response to commensal factors express the chemoattractant CCL-20. 3. CCL-20 recruits CCR-6-expressing lymphocytes comprising B cells, regulatory T cells (Treg) and T cells expressing αβ and γδ T-cell receptors that impact on CAC progression. B cells impact on macrophage polarisation, whereas CCR-6 + γδ T cells provide pathogenic IL-17. 4. IL-6 signalling in αβ T cells releases them from Treg-mediated suppression to exert their effector functions such as T helper (Th)1 and Th17 responses. TAM, tumour-associated macrophage

    Techniques Used: Expressing

    IL-6-polarised M2 macrophages express CCL-20. a Heat map of differently expressed transcripts in tumours derived from 17-week-old HFD-fed Il6r α Fl versus Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol, assessed with a cutoff of a change in expression of 1.5-fold and p -value ≤ 0.025. b IPA analysis diagram of downregulated (green) and upregulated (red) cell recruitment factors in tumour tissues derived from 17-week-old HFD-fed Il6r α Fl versus Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol. Upper number represent respective p -value, lower number fold change. c qPCR analysis of indicated gene expression in tumours of 17-week-old HFD-fed Il6r α Fl versus Il6r α KO ( n = 6–12) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to non-colitic NCD Il6r α Fl colons at day 0. d qPCR analysis of indicated gene expression in distal colons of 8-week-old NCD ( n = 6) and HFD ( n = 7) fed C57BL/6 mice. e qPCR analysis of Ccl20 expression in non-tumour tissue and tumours of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice ( n = 6) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to NCD Il6r α Fl non-tumour at day 62. f Representative immunofluorescent stainings of CCL-20 (red) expression in colons of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol. g qPCR analysis of Ccr6 expression in non-tumour tissue and tumours of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice ( n = 6) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to NCD Il6r α Fl non-tumour at day 62. h qPCR analysis of Ccl20 gene expression in F4/80 MACS-purified macrophages from colitic colons of 10-week-old NCD-fed Il6r α Fl , Il6r α KO and Il6r α myl - KO ( n = 6) at day 13 of the 1.5% AOM/DSS protocol, results are presented relative to macrophages F4/80 MACS sorted from non-colitic Il6r α Fl colons. i CCL-20 produced by M1- and M2-polarised BMDM from control mice ( n = 4) stimulated 24 h with IL-6 or LPS as examined by ELISA. qPCR analysis of Ccl20 gene expression in j M1 ( n = 4–15) or k M2 ( n = 7)-polarised BMDM stimulated with IL-6 or LPS for 8 h from Il6r α Fl and Il6r α KO mice, results are presented relative to LPS-stimulated Il6r α Fl M1. l CCL-20 produced by M2-polarised BMDM stimulated 36 h with LPS from Il6r α Fl and Il6r α KO mice ( n = 6) as examined by ELISA. AOM, azoxymethane; DSS, dextran sodium sulphate; NCD, normal chow diet; HFD, high-fat diet; IPA, Ingenuity pathway analysis; MACS, magnetic-activated cell sorting; BMDM, bone marrow-derived macrophages; n.d., not determined. centre line: median; box limits: 1st and 3rd quartiles; whisker: maximum to minimum, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001 two-tailed unpaired Student’s t -test d , one-way h or two-way ANOVA followed by Fisher LSD c , e , g , i , j , k , l . Scale bar, 50 μm
    Figure Legend Snippet: IL-6-polarised M2 macrophages express CCL-20. a Heat map of differently expressed transcripts in tumours derived from 17-week-old HFD-fed Il6r α Fl versus Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol, assessed with a cutoff of a change in expression of 1.5-fold and p -value ≤ 0.025. b IPA analysis diagram of downregulated (green) and upregulated (red) cell recruitment factors in tumour tissues derived from 17-week-old HFD-fed Il6r α Fl versus Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol. Upper number represent respective p -value, lower number fold change. c qPCR analysis of indicated gene expression in tumours of 17-week-old HFD-fed Il6r α Fl versus Il6r α KO ( n = 6–12) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to non-colitic NCD Il6r α Fl colons at day 0. d qPCR analysis of indicated gene expression in distal colons of 8-week-old NCD ( n = 6) and HFD ( n = 7) fed C57BL/6 mice. e qPCR analysis of Ccl20 expression in non-tumour tissue and tumours of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice ( n = 6) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to NCD Il6r α Fl non-tumour at day 62. f Representative immunofluorescent stainings of CCL-20 (red) expression in colons of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice at day 62 of the 1.5% AOM/DSS protocol. g qPCR analysis of Ccr6 expression in non-tumour tissue and tumours of 17-week-old NCD and HFD-fed Il6r α Fl and Il6r α KO mice ( n = 6) at day 62 of the 1.5% AOM/DSS protocol, results are presented relative to NCD Il6r α Fl non-tumour at day 62. h qPCR analysis of Ccl20 gene expression in F4/80 MACS-purified macrophages from colitic colons of 10-week-old NCD-fed Il6r α Fl , Il6r α KO and Il6r α myl - KO ( n = 6) at day 13 of the 1.5% AOM/DSS protocol, results are presented relative to macrophages F4/80 MACS sorted from non-colitic Il6r α Fl colons. i CCL-20 produced by M1- and M2-polarised BMDM from control mice ( n = 4) stimulated 24 h with IL-6 or LPS as examined by ELISA. qPCR analysis of Ccl20 gene expression in j M1 ( n = 4–15) or k M2 ( n = 7)-polarised BMDM stimulated with IL-6 or LPS for 8 h from Il6r α Fl and Il6r α KO mice, results are presented relative to LPS-stimulated Il6r α Fl M1. l CCL-20 produced by M2-polarised BMDM stimulated 36 h with LPS from Il6r α Fl and Il6r α KO mice ( n = 6) as examined by ELISA. AOM, azoxymethane; DSS, dextran sodium sulphate; NCD, normal chow diet; HFD, high-fat diet; IPA, Ingenuity pathway analysis; MACS, magnetic-activated cell sorting; BMDM, bone marrow-derived macrophages; n.d., not determined. centre line: median; box limits: 1st and 3rd quartiles; whisker: maximum to minimum, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001 two-tailed unpaired Student’s t -test d , one-way h or two-way ANOVA followed by Fisher LSD c , e , g , i , j , k , l . Scale bar, 50 μm

    Techniques Used: Derivative Assay, Mouse Assay, Expressing, Indirect Immunoperoxidase Assay, Real-time Polymerase Chain Reaction, Magnetic Cell Separation, Purification, Produced, Enzyme-linked Immunosorbent Assay, FACS, Whisker Assay, Two Tailed Test

    11) Product Images from "Improved Insulin Sensitivity in High Fat- and High Cholesterol-fed Ldlr−/− Mice with Macrophage-specific Transgenic Expression of Cholesteryl Ester Hydrolase"

    Article Title: Improved Insulin Sensitivity in High Fat- and High Cholesterol-fed Ldlr−/− Mice with Macrophage-specific Transgenic Expression of Cholesteryl Ester Hydrolase

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.069781

    CEH overexpression decreases the proinflammatory cytokine production from macrophages. A , thioglycollate-elicited macrophages were obtained from Ldlr −/− and Ldlr −/− CEHTg mice and lipid loaded with acetylated low density lipoprotein ( AcLDL ; 50 μg/ml). IL-1β and MCP-1 levels in the conditioned medium were determined by ELISA. Data are expressed as mean ± S.D. ( n = 3). B , thioglycollate-elicited macrophages were obtained from Western diet-fed Ldlr −/− and Ldlr −/− CEHTg mice, and total RNA was isolated from adherent cells 2 h after plating. Expression of IL-1β, IL-6, and MCP-1 was determined by real time reverse transcription-PCR, and data (mean ± S.D.; n = 3) are expressed as percent mRNA levels in Ldlr −/− CEHTg mice. *, p
    Figure Legend Snippet: CEH overexpression decreases the proinflammatory cytokine production from macrophages. A , thioglycollate-elicited macrophages were obtained from Ldlr −/− and Ldlr −/− CEHTg mice and lipid loaded with acetylated low density lipoprotein ( AcLDL ; 50 μg/ml). IL-1β and MCP-1 levels in the conditioned medium were determined by ELISA. Data are expressed as mean ± S.D. ( n = 3). B , thioglycollate-elicited macrophages were obtained from Western diet-fed Ldlr −/− and Ldlr −/− CEHTg mice, and total RNA was isolated from adherent cells 2 h after plating. Expression of IL-1β, IL-6, and MCP-1 was determined by real time reverse transcription-PCR, and data (mean ± S.D.; n = 3) are expressed as percent mRNA levels in Ldlr −/− CEHTg mice. *, p

    Techniques Used: Over Expression, Mouse Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Isolation, Expressing, Polymerase Chain Reaction

    12) Product Images from "The role of TRPV1 in the CD4+ T cell-mediated inflammatory response of allergic rhinitis"

    Article Title: The role of TRPV1 in the CD4+ T cell-mediated inflammatory response of allergic rhinitis

    Journal: Oncotarget

    doi:

    Expression of TRPV1 and its regulation of cytokine production and T cell receptor signaling in CD4 + T lymphocytes A. The expression of TRPV1 protein in the CD4 + T cells was analyzed by FACS and confocal microscopic analyses. B. Protein concentrations of the cytokines IL-4, IL-5, IL-6, IL-10, IL-17, and IFN-γ secreted by CD4 + T cells isolated from TRPV1 (+/+) and TRPV1 (−/−) mice splenocytes after stimulation with αCD3/CD28. C. The immunoblotting results of T cell recptor signaling pathways in CD4 + T cells isolated from TRPV1 (+/+) and TRPV1 (−/−) mice. Phospho-p65 represents NF-κB activity and phospho-p38, phospho-ERK and phospho-JNK represents MAPK pathway. D. The secreted protein concentrations of the cytokines IL-4, IL-5, IL-6, IL-10, IL-17, and IFN-γ in the culture media of human Jurkat T cells pre-incubated with BCTC after stimulation with αCD3/CD28 were determined by ELISA. E. T cell receptor signaling pathways in human Jurkat T cells with or without preincubation of TRPV1 inhibitor, BCTC. TCR was activated by anti-CD3/CD28 and two different concentration of BCTC was used. The statistical P values are presented as * ( P
    Figure Legend Snippet: Expression of TRPV1 and its regulation of cytokine production and T cell receptor signaling in CD4 + T lymphocytes A. The expression of TRPV1 protein in the CD4 + T cells was analyzed by FACS and confocal microscopic analyses. B. Protein concentrations of the cytokines IL-4, IL-5, IL-6, IL-10, IL-17, and IFN-γ secreted by CD4 + T cells isolated from TRPV1 (+/+) and TRPV1 (−/−) mice splenocytes after stimulation with αCD3/CD28. C. The immunoblotting results of T cell recptor signaling pathways in CD4 + T cells isolated from TRPV1 (+/+) and TRPV1 (−/−) mice. Phospho-p65 represents NF-κB activity and phospho-p38, phospho-ERK and phospho-JNK represents MAPK pathway. D. The secreted protein concentrations of the cytokines IL-4, IL-5, IL-6, IL-10, IL-17, and IFN-γ in the culture media of human Jurkat T cells pre-incubated with BCTC after stimulation with αCD3/CD28 were determined by ELISA. E. T cell receptor signaling pathways in human Jurkat T cells with or without preincubation of TRPV1 inhibitor, BCTC. TCR was activated by anti-CD3/CD28 and two different concentration of BCTC was used. The statistical P values are presented as * ( P

    Techniques Used: Expressing, FACS, Isolation, Mouse Assay, Activity Assay, Incubation, Enzyme-linked Immunosorbent Assay, Concentration Assay

    13) Product Images from "Mechanism and Treatment of Renal Fibrosis: Moderate aging does not exacerbate cisplatin-induced kidney injury or fibrosis despite altered inflammatory cytokine expression and immune cell infiltration"

    Article Title: Mechanism and Treatment of Renal Fibrosis: Moderate aging does not exacerbate cisplatin-induced kidney injury or fibrosis despite altered inflammatory cytokine expression and immune cell infiltration

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00463.2018

    Changes in inflammatory cytokine and chemokine levels with the standard-dosing regimen of cisplatin (0 or 25 mg/kg once) in 8- and 40-wk-old male FVB mice. Tumor necrosis factor-α ( Tnfa ; A ), interleukin-6 ( Il6 ; B ), chemokine (C-X-C motif) ligand 1 ( Cxcl1 ; C ), and monocyte chemoattractant protein 1 ( Mcp1 ; D ) were measured in the kidney cortex by real-time quantitative RT-PCR. Values are means ± SE; n = 5–10. * P
    Figure Legend Snippet: Changes in inflammatory cytokine and chemokine levels with the standard-dosing regimen of cisplatin (0 or 25 mg/kg once) in 8- and 40-wk-old male FVB mice. Tumor necrosis factor-α ( Tnfa ; A ), interleukin-6 ( Il6 ; B ), chemokine (C-X-C motif) ligand 1 ( Cxcl1 ; C ), and monocyte chemoattractant protein 1 ( Mcp1 ; D ) were measured in the kidney cortex by real-time quantitative RT-PCR. Values are means ± SE; n = 5–10. * P

    Techniques Used: Mouse Assay, Quantitative RT-PCR

    Changes in inflammatory cytokine and chemokine levels with the repeated-dosing regimen of cisplatin (0 or 7 mg/kg once a week for 4 wk) in 8- and 40-wk-old male FVB mice. Tumor necrosis factor-α ( Tnfa ; A ), interleukin-6 ( Il6 ; B ), chemokine (C-X-C motif) ligand 1 ( Cxcl1 ; C ), and monocyte chemoattractant protein 1 ( Mcp1 ; D ) were measured in the kidney cortex by real-time quantitative RT-PCR. Values are means ± SE; n = 5–10. ** P
    Figure Legend Snippet: Changes in inflammatory cytokine and chemokine levels with the repeated-dosing regimen of cisplatin (0 or 7 mg/kg once a week for 4 wk) in 8- and 40-wk-old male FVB mice. Tumor necrosis factor-α ( Tnfa ; A ), interleukin-6 ( Il6 ; B ), chemokine (C-X-C motif) ligand 1 ( Cxcl1 ; C ), and monocyte chemoattractant protein 1 ( Mcp1 ; D ) were measured in the kidney cortex by real-time quantitative RT-PCR. Values are means ± SE; n = 5–10. ** P

    Techniques Used: Mouse Assay, Quantitative RT-PCR

    14) Product Images from "Stromal senescence establishes an immunosuppressive microenvironment that drives tumorigenesis"

    Article Title: Stromal senescence establishes an immunosuppressive microenvironment that drives tumorigenesis

    Journal: Nature Communications

    doi: 10.1038/ncomms11762

    Senescence-derived IL-6 is necessary to drive G-MDSC accumulation and tumour promotion. ( a ) Senescence-associated β-galactosidase (SA-βgal) staining (blue) of shRFP-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼65% of cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( b ) SA-βgal staining (blue) of shIL6#1-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼55% of shIL6 #1 cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( c ) SA-βgal staining (blue) of shIL6#2-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼55% of shIL6 #2 cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( d ) Quantification of SA-βgal-positive shRFP-, shIL6 #1-, and shIL6 #2-expressing cells shown in a – c . Data are mean %-positive cells+s.e.m. * P value
    Figure Legend Snippet: Senescence-derived IL-6 is necessary to drive G-MDSC accumulation and tumour promotion. ( a ) Senescence-associated β-galactosidase (SA-βgal) staining (blue) of shRFP-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼65% of cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( b ) SA-βgal staining (blue) of shIL6#1-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼55% of shIL6 #1 cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( c ) SA-βgal staining (blue) of shIL6#2-expressing MSFs treated with 10 μM TAM (Sen) or vehicle (Non-Sen) revealed ∼55% of shIL6 #2 cells were senescent following TAM treatment. Scale bar, 100 μm. n =3. ( d ) Quantification of SA-βgal-positive shRFP-, shIL6 #1-, and shIL6 #2-expressing cells shown in a – c . Data are mean %-positive cells+s.e.m. * P value

    Techniques Used: Derivative Assay, Staining, Expressing

    IL-6 within senescent microenvironments mediates immunosuppression and tumour promotion. ( a ) Flow cytometry for G-MDSC following IL-6 depletion in non-senescent (Non-Sen) or senescent (Sen) isografts. * P value
    Figure Legend Snippet: IL-6 within senescent microenvironments mediates immunosuppression and tumour promotion. ( a ) Flow cytometry for G-MDSC following IL-6 depletion in non-senescent (Non-Sen) or senescent (Sen) isografts. * P value

    Techniques Used: Flow Cytometry, Cytometry

    15) Product Images from "Downregulation of macrophage Irs2 by hyperinsulinemia impairs IL-4-indeuced M2a-subtype macrophage activation in obesity"

    Article Title: Downregulation of macrophage Irs2 by hyperinsulinemia impairs IL-4-indeuced M2a-subtype macrophage activation in obesity

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07358-9

    The Irs2 mRNA levels and number of M2a-subtype MΦs were elevated in HF diet-fed M IR KO mice. a Expression levels of IR , IL-4R , Irs2 and STAT6 in the siglecF - CD11b + F4/80 + cells of the SVF of the adipose tissue from the control and M IR KO mice ( n = 4–6). b IL-4-induced STAT6, Irs2 and Akt phosphorylation in peritoneal MΦs of HF diet-fed M IR KO mice ( n = 3–6). c Irs2 expression levels in the BMDM of the control and M IR KO mice after 100 nM insulin stimulation for 3 h ( n = 3–4). d IL-4-induced M2a-subtype marker genes in the BMDM after 100 nM insulin pretreatment for 8 h ( n = 3–4). e IL-4-induced Arg1 expression levels in the BMDM pretreated with 100 nM insulin for 8 h after siFoxO1, siNCoR1, or siHDAC3 treatment ( n = 4–16). f MCP-1 , CCR2 , IL-18 and IL-6 expression levels in 3T3-L1 cells in co-culture with BMDM of the C57BL/6 mice and 3T3-L1 cells after insulin stimulation for 24 h ( n = 7). The data are mean ± SEM. followed by one-way ANOVA with a post hoc test or Student’s t test. * P
    Figure Legend Snippet: The Irs2 mRNA levels and number of M2a-subtype MΦs were elevated in HF diet-fed M IR KO mice. a Expression levels of IR , IL-4R , Irs2 and STAT6 in the siglecF - CD11b + F4/80 + cells of the SVF of the adipose tissue from the control and M IR KO mice ( n = 4–6). b IL-4-induced STAT6, Irs2 and Akt phosphorylation in peritoneal MΦs of HF diet-fed M IR KO mice ( n = 3–6). c Irs2 expression levels in the BMDM of the control and M IR KO mice after 100 nM insulin stimulation for 3 h ( n = 3–4). d IL-4-induced M2a-subtype marker genes in the BMDM after 100 nM insulin pretreatment for 8 h ( n = 3–4). e IL-4-induced Arg1 expression levels in the BMDM pretreated with 100 nM insulin for 8 h after siFoxO1, siNCoR1, or siHDAC3 treatment ( n = 4–16). f MCP-1 , CCR2 , IL-18 and IL-6 expression levels in 3T3-L1 cells in co-culture with BMDM of the C57BL/6 mice and 3T3-L1 cells after insulin stimulation for 24 h ( n = 7). The data are mean ± SEM. followed by one-way ANOVA with a post hoc test or Student’s t test. * P

    Techniques Used: Mouse Assay, Expressing, Marker, Co-Culture Assay

    IL-4-induced M2a-subtype MΦ activation was impaired in WAT of the HF diet-fed M Irs2 KO mice. a The percentages of siglecF - CD11b + F4/80 + , siglecF - CD11b + F4/80 + CD11c + CD206 - (M1-type MΦs) and siglecF - CD11b + F4/80 + CD11c - CD206 + (M2-type MΦs) cells in the SVF of the adipose tissue ( n = 10–14). b Expression levels of M2-type and M1-type MΦ marker genes in the siglecF - CD11b + F4/80 + cells in the SVF of the adipose tissue from the control and M Irs2 KO mice ( n = 10–14). c Expression levels of M2a-subtype MΦ marker genes in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation for 48 h ( n = 6–8). d Arginase activity in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation for 24 h ( n = 7–10). e Expression levels of the M1-type MΦ marker genes in the BMDM of the control and M Irs2 KO mice after LPS stimulation for 48 h ( n = 3). f MCP-1 , CCR2 , IL-18 and IL-6 expression levels in 3T3-L1 cells in co-culture with the BMDM of M Irs2 KO mice ( n = 12). g Irs2 phosphorylation and protein levels in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation ( n = 3–4). h Akt phosphorylation and protein levels in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without LY294002 treatment ( n = 3). i IL-4-induced Akt phosphorylation in the BMDM of the M Irs2 KO mice ( n = 4). j Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without LY294002 treatment ( n = 4–6). k Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without CA-FoxO1 treatment ( n = 4–6). l FoxO1 phosphorylation and protein levels in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation ( n = 3–4). m Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the M Irs2 KO mice after IL-4 stimulation with siFoxO1 transfection ( n = 5–6). The data are mean ± SEM. followed by one-way ANOVA with a post hoc test or Student’s t test. * P
    Figure Legend Snippet: IL-4-induced M2a-subtype MΦ activation was impaired in WAT of the HF diet-fed M Irs2 KO mice. a The percentages of siglecF - CD11b + F4/80 + , siglecF - CD11b + F4/80 + CD11c + CD206 - (M1-type MΦs) and siglecF - CD11b + F4/80 + CD11c - CD206 + (M2-type MΦs) cells in the SVF of the adipose tissue ( n = 10–14). b Expression levels of M2-type and M1-type MΦ marker genes in the siglecF - CD11b + F4/80 + cells in the SVF of the adipose tissue from the control and M Irs2 KO mice ( n = 10–14). c Expression levels of M2a-subtype MΦ marker genes in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation for 48 h ( n = 6–8). d Arginase activity in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation for 24 h ( n = 7–10). e Expression levels of the M1-type MΦ marker genes in the BMDM of the control and M Irs2 KO mice after LPS stimulation for 48 h ( n = 3). f MCP-1 , CCR2 , IL-18 and IL-6 expression levels in 3T3-L1 cells in co-culture with the BMDM of M Irs2 KO mice ( n = 12). g Irs2 phosphorylation and protein levels in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation ( n = 3–4). h Akt phosphorylation and protein levels in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without LY294002 treatment ( n = 3). i IL-4-induced Akt phosphorylation in the BMDM of the M Irs2 KO mice ( n = 4). j Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without LY294002 treatment ( n = 4–6). k Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the C57BL/6 mice after IL-4 stimulation with or without CA-FoxO1 treatment ( n = 4–6). l FoxO1 phosphorylation and protein levels in the BMDM of the control and M Irs2 KO mice after IL-4 stimulation ( n = 3–4). m Expression levels of the M2a-subtype MΦ marker genes in the BMDM of the M Irs2 KO mice after IL-4 stimulation with siFoxO1 transfection ( n = 5–6). The data are mean ± SEM. followed by one-way ANOVA with a post hoc test or Student’s t test. * P

    Techniques Used: Activation Assay, Mouse Assay, Expressing, Marker, Activity Assay, Co-Culture Assay, Transfection

    16) Product Images from "Strain-Specific Differences in House Dust Mite (Dermatophagoides farinae)-Induced Mouse Models of Allergic Rhinitis"

    Article Title: Strain-Specific Differences in House Dust Mite (Dermatophagoides farinae)-Induced Mouse Models of Allergic Rhinitis

    Journal: Clinical and Experimental Otorhinolaryngology

    doi: 10.21053/ceo.2019.01837

    Nasal mRNA expression of cytokines. Shown are interleukin (IL)-4 (A), IL-5 (B), IL-6 (C), IL-10 (D), IL-17 (E), and interferon (IFN)-γ (F) levels in the control group of BALB/c mice (group A), BALB/c mice exposed to 25 µg of Dermatophagoides farinae (Der f1; group B), BALB/c mice group exposed to 100 µg of Der f1 (group C), control group of C57BL/6 mice (group D), C57BL/6 mice exposed to 25 µg of Der f1 (group E), and C57BL/6 mice exposed to 100 µg of Der f1 (group F). Bars represent standard error. Significantly different from control mice or significantly different between strains, *** P
    Figure Legend Snippet: Nasal mRNA expression of cytokines. Shown are interleukin (IL)-4 (A), IL-5 (B), IL-6 (C), IL-10 (D), IL-17 (E), and interferon (IFN)-γ (F) levels in the control group of BALB/c mice (group A), BALB/c mice exposed to 25 µg of Dermatophagoides farinae (Der f1; group B), BALB/c mice group exposed to 100 µg of Der f1 (group C), control group of C57BL/6 mice (group D), C57BL/6 mice exposed to 25 µg of Der f1 (group E), and C57BL/6 mice exposed to 100 µg of Der f1 (group F). Bars represent standard error. Significantly different from control mice or significantly different between strains, *** P

    Techniques Used: Expressing, Mouse Assay

    Cytokine levels from splenocyte culture; interleukin (IL)-4 (A), IL-5 (B), IL-6 (C), IL-10 (D), IL-17 (E) and interferon (IFN)-γ (F) in the control group of BALB/c mice (group A), BALB/c mice exposed to 25 µg of Dermatophagoides farinae (Der f1; group B), BALB/c mice group exposed to 100 µg of Der f1 (group C), control group of C57BL/6 mice (group D), C57BL/6 mice exposed to 25 µg of Der f1 (group E), and C57BL/6 mice exposed to 100 µg of Der f1 (group F). Bars represent standard error. Significantly different from control mice or significantly different between strains, * P
    Figure Legend Snippet: Cytokine levels from splenocyte culture; interleukin (IL)-4 (A), IL-5 (B), IL-6 (C), IL-10 (D), IL-17 (E) and interferon (IFN)-γ (F) in the control group of BALB/c mice (group A), BALB/c mice exposed to 25 µg of Dermatophagoides farinae (Der f1; group B), BALB/c mice group exposed to 100 µg of Der f1 (group C), control group of C57BL/6 mice (group D), C57BL/6 mice exposed to 25 µg of Der f1 (group E), and C57BL/6 mice exposed to 100 µg of Der f1 (group F). Bars represent standard error. Significantly different from control mice or significantly different between strains, * P

    Techniques Used: Mouse Assay

    17) Product Images from "Berberine Protects against Neuronal Damage via Suppression of Glia-Mediated Inflammation in Traumatic Brain Injury"

    Article Title: Berberine Protects against Neuronal Damage via Suppression of Glia-Mediated Inflammation in Traumatic Brain Injury

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0115694

    Berberine inhibited IL-1β-induced activation of TLR4/MyD88/NF-κB signaling in mixed glia and attenuated IL-1β-induced microglial activation. Representative immunoblots showing that co-treatment of 50 µM berberine with IL-1β for 1 day significantly reduced IL-1β-induced ( A ) toll-like receptor 4 (TLR4) expression ( B ) adapter protein myeloid differentiation factor 88 (MyD88) expression ( C ) p65 nuclear translocation ( D ) inducible nitric oxide synthase (iNOS) expression and ( E ) cyclooxygenase-2 (COX-2) expression by primary mixed glia. ( F, G ) Co-treatment of 50 µM berberine with IL-1β for 1 day significantly reduced IL-1β-induced secretion of IL-6 and macrophage inflammatory protein (MIP-2) as assessed by enzyme-linked immunosorbent assay from the supernatant of mixed glial cultures. Co-treatment of 50 µM berberine with IL-1β for 2 days significantly attenuated IL-1β-induced release of ( H ) nitric oxide (NO), ( I ) IL-6 and ( J ) MIP-2 from the supernatant of primary microglial cultures. Values are presented as mean ± SEM of four independent experiments. * P
    Figure Legend Snippet: Berberine inhibited IL-1β-induced activation of TLR4/MyD88/NF-κB signaling in mixed glia and attenuated IL-1β-induced microglial activation. Representative immunoblots showing that co-treatment of 50 µM berberine with IL-1β for 1 day significantly reduced IL-1β-induced ( A ) toll-like receptor 4 (TLR4) expression ( B ) adapter protein myeloid differentiation factor 88 (MyD88) expression ( C ) p65 nuclear translocation ( D ) inducible nitric oxide synthase (iNOS) expression and ( E ) cyclooxygenase-2 (COX-2) expression by primary mixed glia. ( F, G ) Co-treatment of 50 µM berberine with IL-1β for 1 day significantly reduced IL-1β-induced secretion of IL-6 and macrophage inflammatory protein (MIP-2) as assessed by enzyme-linked immunosorbent assay from the supernatant of mixed glial cultures. Co-treatment of 50 µM berberine with IL-1β for 2 days significantly attenuated IL-1β-induced release of ( H ) nitric oxide (NO), ( I ) IL-6 and ( J ) MIP-2 from the supernatant of primary microglial cultures. Values are presented as mean ± SEM of four independent experiments. * P

    Techniques Used: Activation Assay, Western Blot, Expressing, Translocation Assay, Enzyme-linked Immunosorbent Assay

    Delayed berberine treatment attenuated contusion volume, neuronal death and apoptosis and reduced inflammation after TBI. Treatment with 10 mg·kg −1 s berberine at 3 h post-injury significantly reduced ( A ) contusion volume as assessed by cresyl violet staining, ( B ) the number of Fluoro-Jade B (FJB)-positive neurons in the cortical contusion margin and ( C ) the cleaved caspase-3 (cCP-3) level in the ipsilateral hemisphere than vehicle-treated mice at 1 day post-TBI. The total number of FJB-positive cells is expressed as the mean number per field of view (0.8 mm 2 ). ( D ) Bar graphs of IL-1β, IL-6, MCP-1, and MIP-2 protein concentrations, as assessed by enzyme-linked immunosorbent assays (ELISA) in the ipsilateral cortices of sham control, vehicle-treated, and 10 mg·kg −1 berberine-treated mice at 1 day post-injury. Treatment with 10 mg·kg −1 s berberine at 3 h post-injury significantly reduced IL-1β, IL-6, MCP-1, and MIP-2 protein levels compared with vehicle-treated mice. Values are presented as means ± SEM; *** P
    Figure Legend Snippet: Delayed berberine treatment attenuated contusion volume, neuronal death and apoptosis and reduced inflammation after TBI. Treatment with 10 mg·kg −1 s berberine at 3 h post-injury significantly reduced ( A ) contusion volume as assessed by cresyl violet staining, ( B ) the number of Fluoro-Jade B (FJB)-positive neurons in the cortical contusion margin and ( C ) the cleaved caspase-3 (cCP-3) level in the ipsilateral hemisphere than vehicle-treated mice at 1 day post-TBI. The total number of FJB-positive cells is expressed as the mean number per field of view (0.8 mm 2 ). ( D ) Bar graphs of IL-1β, IL-6, MCP-1, and MIP-2 protein concentrations, as assessed by enzyme-linked immunosorbent assays (ELISA) in the ipsilateral cortices of sham control, vehicle-treated, and 10 mg·kg −1 berberine-treated mice at 1 day post-injury. Treatment with 10 mg·kg −1 s berberine at 3 h post-injury significantly reduced IL-1β, IL-6, MCP-1, and MIP-2 protein levels compared with vehicle-treated mice. Values are presented as means ± SEM; *** P

    Techniques Used: Staining, Mouse Assay, Enzyme-linked Immunosorbent Assay

    18) Product Images from "Natural Killer Cells from the Subcutaneous Adipose Tissue Underexpress the NKp30 and NKp44 in Obese Persons and Are Less Active against Major Histocompatibility Complex Class I Non-Expressing Neoplastic Cells"

    Article Title: Natural Killer Cells from the Subcutaneous Adipose Tissue Underexpress the NKp30 and NKp44 in Obese Persons and Are Less Active against Major Histocompatibility Complex Class I Non-Expressing Neoplastic Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01486

    The gene expression profile of IL-1b, IL-6, IL-8, IL-10, IL-12a, IL-15, interferon gamma (IFNγ), and tumor necrosis factor alpha (TNFα) in subcutaneous adipose tissue-derived (A) and peripheral blood (PB) (B) of total natural killer cells (NKs) in obese and lean cases via qRT-PCR. The levels of these cytokines in obese persons are compared to the PB total NK cells of lean cases as the control. PBL, peripheral blood of lean cases; PBO, peripheral blood of obese cases; SATO, subcutaneous adipose tissue derived of obese cases.
    Figure Legend Snippet: The gene expression profile of IL-1b, IL-6, IL-8, IL-10, IL-12a, IL-15, interferon gamma (IFNγ), and tumor necrosis factor alpha (TNFα) in subcutaneous adipose tissue-derived (A) and peripheral blood (PB) (B) of total natural killer cells (NKs) in obese and lean cases via qRT-PCR. The levels of these cytokines in obese persons are compared to the PB total NK cells of lean cases as the control. PBL, peripheral blood of lean cases; PBO, peripheral blood of obese cases; SATO, subcutaneous adipose tissue derived of obese cases.

    Techniques Used: Expressing, Derivative Assay, Quantitative RT-PCR

    19) Product Images from "Cytokine treatment optimises the immunotherapeutic effects of umbilical cord-derived MSC for treatment of inflammatory liver disease"

    Article Title: Cytokine treatment optimises the immunotherapeutic effects of umbilical cord-derived MSC for treatment of inflammatory liver disease

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0590-6

    Effect of ucMSC on ex vivo liver slices. Gene expression levels and cytokine/chemokine levels were measured in liver slices treated with LPS and supernatant, respectively. ( a )( b )( c ) Both gene expression levels and secretion levels were measured for MCP-1, TNF-α and IL-6. In addition, d gene expression levels of IP-10 and also e CXCL1 were measured in the medium. Results are shown as means ± SEM (n = 6). * Indicates significant difference ( p
    Figure Legend Snippet: Effect of ucMSC on ex vivo liver slices. Gene expression levels and cytokine/chemokine levels were measured in liver slices treated with LPS and supernatant, respectively. ( a )( b )( c ) Both gene expression levels and secretion levels were measured for MCP-1, TNF-α and IL-6. In addition, d gene expression levels of IP-10 and also e CXCL1 were measured in the medium. Results are shown as means ± SEM (n = 6). * Indicates significant difference ( p

    Techniques Used: Ex Vivo, Expressing

    20) Product Images from "Interleukin-6: A Constitutive Modulator of Glycoprotein 130, Neuroinflammatory and Cell Survival Signaling in Retina"

    Article Title: Interleukin-6: A Constitutive Modulator of Glycoprotein 130, Neuroinflammatory and Cell Survival Signaling in Retina

    Journal: Journal of clinical & cellular immunology

    doi: 10.4172/2155-9899.1000439

    IL-6 deficiency leads to decreased gp130 protein in the retina. ( A ) Fold difference of Gp130 mRNA in the IL-6−/− retina normalized to WT Gp130 mRNA (dotted line) via the ΔΔCt method. ( B ) Densitometry analysis (right) of gp130 and β-actin immunoblotting (left) in WT (white) and IL-6−/− (gray) retina.; student’s t-test. Error bars indicates STDEV. ( C–D ) Whole mount ( C ) and sagittal cross sections ( D ) of WT (left) and IL-6−/− (right) retina reveal co-localization (arrowheads; C and arrows; D) of gp130 immunolabeling (green) with β-Tubulin+ RGCs. Error bars represent standard deviation and asterisks indicate p
    Figure Legend Snippet: IL-6 deficiency leads to decreased gp130 protein in the retina. ( A ) Fold difference of Gp130 mRNA in the IL-6−/− retina normalized to WT Gp130 mRNA (dotted line) via the ΔΔCt method. ( B ) Densitometry analysis (right) of gp130 and β-actin immunoblotting (left) in WT (white) and IL-6−/− (gray) retina.; student’s t-test. Error bars indicates STDEV. ( C–D ) Whole mount ( C ) and sagittal cross sections ( D ) of WT (left) and IL-6−/− (right) retina reveal co-localization (arrowheads; C and arrows; D) of gp130 immunolabeling (green) with β-Tubulin+ RGCs. Error bars represent standard deviation and asterisks indicate p

    Techniques Used: Immunolabeling, Standard Deviation

    IL-6 deficiency influences glaucoma-related changes in gene expression of neuroinflammory, cell health and gp130 modulators. ( A–E ) Percent gene expression of ( A ) Tnfα , Il-β ( B ) Bcl-xl ( C ), Bax and ( D ) Socs3 ( E ) mRNA in either WT or IL-6−/− eyes injected with saline (white) or microbeads (gray). mRNA levels were normalized to respective naive expression levels (dotted line) via the ΔΔCt method. Statistical significance (p
    Figure Legend Snippet: IL-6 deficiency influences glaucoma-related changes in gene expression of neuroinflammory, cell health and gp130 modulators. ( A–E ) Percent gene expression of ( A ) Tnfα , Il-β ( B ) Bcl-xl ( C ), Bax and ( D ) Socs3 ( E ) mRNA in either WT or IL-6−/− eyes injected with saline (white) or microbeads (gray). mRNA levels were normalized to respective naive expression levels (dotted line) via the ΔΔCt method. Statistical significance (p

    Techniques Used: Expressing, Injection

    Review of IL-6 signaling. Under naïve and pathological conditions, IL-6 binds to IL-6Rα, followed by recruitment of signal transducer gp130 (purple arrows). Upon formation of the IL-6/IL-6Rα/gp130 complex, gp130 phosphorylates Jak2, followed by subsequent activation/phosphorylation of STAT3. Phosphorylated STAT3 dimerizes and translocates to the nucleus to drive transcription of genes associated with cell survival (Bxl-XL, BCL-2) and gp130 inhibitory feedback (Socs3). Socs3 can also act to inhibit NFκB, which regulates the expression of inflammatory cytokines (i.e. IL-6, IL-1β, TNFα).
    Figure Legend Snippet: Review of IL-6 signaling. Under naïve and pathological conditions, IL-6 binds to IL-6Rα, followed by recruitment of signal transducer gp130 (purple arrows). Upon formation of the IL-6/IL-6Rα/gp130 complex, gp130 phosphorylates Jak2, followed by subsequent activation/phosphorylation of STAT3. Phosphorylated STAT3 dimerizes and translocates to the nucleus to drive transcription of genes associated with cell survival (Bxl-XL, BCL-2) and gp130 inhibitory feedback (Socs3). Socs3 can also act to inhibit NFκB, which regulates the expression of inflammatory cytokines (i.e. IL-6, IL-1β, TNFα).

    Techniques Used: Activation Assay, Activated Clotting Time Assay, Expressing

    IL-6 deficiency alters baseline gene expression for neuroinflammation, cell health and gp130 regulatory signaling. Percent gene expression of ( A ) Tnfα , Il-β ( B ) Bcl-xl , Bax and ( C ) Socs3 mRNA in the IL-6−/− retina normalized to WT expression levels (dotted line) via the ΔΔCt method. Asterisks indicate p
    Figure Legend Snippet: IL-6 deficiency alters baseline gene expression for neuroinflammation, cell health and gp130 regulatory signaling. Percent gene expression of ( A ) Tnfα , Il-β ( B ) Bcl-xl , Bax and ( C ) Socs3 mRNA in the IL-6−/− retina normalized to WT expression levels (dotted line) via the ΔΔCt method. Asterisks indicate p

    Techniques Used: Expressing

    IL-6 deficiency does not impact microbead-induced IOP elevation. Average IOP of eyes injected with either saline (white) or microbeads (gray) over 4 weeks. Error bars represent standard error of the mean and asterisks indicate p
    Figure Legend Snippet: IL-6 deficiency does not impact microbead-induced IOP elevation. Average IOP of eyes injected with either saline (white) or microbeads (gray) over 4 weeks. Error bars represent standard error of the mean and asterisks indicate p

    Techniques Used: Injection

    21) Product Images from "Sex- and cell-dependent contribution of peripheral high mobility group box 1 and TLR4 in arthritis-induced pain"

    Article Title: Sex- and cell-dependent contribution of peripheral high mobility group box 1 and TLR4 in arthritis-induced pain

    Journal: Pain

    doi: 10.1097/j.pain.0000000000002034

    Intra-articular injection of disulfide HMGB1, but not all-thiol HMGB1, induces mRNA levels of inflammatory factors at different timepoints in ankle joints of male and female mice. (A) mRNA expression for HMGB1 receptors Tlr2 , Tlr4 , and Rage in noninjected ankle joints of males and female mice. mRNA expression for inflammatory factors (B) Tnf , (C), Il1b, (D) Il6 , (E) Ccl2 , (F) Cxcl1 , (G) Cxcl2 , (H) Cox2 , and (I) Ngf in male and female mice injected with dsHMG or atHMG. Phosphate buffered saline-injected mice were used as vehicle control group and depicted as dashed lines in the graphs. Data are presented as mean ± SEM, n = 4 to 8 mice/group, * P
    Figure Legend Snippet: Intra-articular injection of disulfide HMGB1, but not all-thiol HMGB1, induces mRNA levels of inflammatory factors at different timepoints in ankle joints of male and female mice. (A) mRNA expression for HMGB1 receptors Tlr2 , Tlr4 , and Rage in noninjected ankle joints of males and female mice. mRNA expression for inflammatory factors (B) Tnf , (C), Il1b, (D) Il6 , (E) Ccl2 , (F) Cxcl1 , (G) Cxcl2 , (H) Cox2 , and (I) Ngf in male and female mice injected with dsHMG or atHMG. Phosphate buffered saline-injected mice were used as vehicle control group and depicted as dashed lines in the graphs. Data are presented as mean ± SEM, n = 4 to 8 mice/group, * P

    Techniques Used: Injection, Mouse Assay, Expressing

    Addition of disulfide HMGB1 in culture induces higher levels of proinflammatory factors in male compared to female macrophages. Levels of (A) TNF, (B) IL-6, (C) CCL2, and (D) CXCL1 measured in culture supernatant after 24-hour stimulation of 1 μg/mL disulfide HMGB1 or phosphate buffered saline (vehicle control) of macrophages generated from male and female mice. Data are presented as mean ± SEM, n = 4 mice/group, * P
    Figure Legend Snippet: Addition of disulfide HMGB1 in culture induces higher levels of proinflammatory factors in male compared to female macrophages. Levels of (A) TNF, (B) IL-6, (C) CCL2, and (D) CXCL1 measured in culture supernatant after 24-hour stimulation of 1 μg/mL disulfide HMGB1 or phosphate buffered saline (vehicle control) of macrophages generated from male and female mice. Data are presented as mean ± SEM, n = 4 mice/group, * P

    Techniques Used: Generated, Mouse Assay

    22) Product Images from "Adipose Tissue-Derived Mesenchymal Stem Cells Increase Skin Allograft Survival and Inhibit Th-17 Immune Response"

    Article Title: Adipose Tissue-Derived Mesenchymal Stem Cells Increase Skin Allograft Survival and Inhibit Th-17 Immune Response

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0076396

    ADSC change the cytokine milieu in vivo and block Th-17 responses. C57BL/6 mice were grafted with full thickness allogeneic tail skin from CBA/J mice and treated or not with donor (CBA/J) ADSC. Tissues were analyzed on days 3 and 10 after transplantation. RNA was isolated from (A) draining axillary lymph nodes and (B) skin. Gene expression of Foxp3, TGF-β, IL-10, IFN-γ, IL-17 and IL-6 was assessed by quantitative RT-PCR. Samples were normalized by expression of an endogenous housekeeping gene (HPRT). Data are represented as mean ± SEM; n = 3 independent experiments done in triplicate, leading to a total of ≥10 independent values for each point. *P
    Figure Legend Snippet: ADSC change the cytokine milieu in vivo and block Th-17 responses. C57BL/6 mice were grafted with full thickness allogeneic tail skin from CBA/J mice and treated or not with donor (CBA/J) ADSC. Tissues were analyzed on days 3 and 10 after transplantation. RNA was isolated from (A) draining axillary lymph nodes and (B) skin. Gene expression of Foxp3, TGF-β, IL-10, IFN-γ, IL-17 and IL-6 was assessed by quantitative RT-PCR. Samples were normalized by expression of an endogenous housekeeping gene (HPRT). Data are represented as mean ± SEM; n = 3 independent experiments done in triplicate, leading to a total of ≥10 independent values for each point. *P

    Techniques Used: In Vivo, Blocking Assay, Mouse Assay, Crocin Bleaching Assay, Transplantation Assay, Isolation, Expressing, Quantitative RT-PCR

    23) Product Images from "Lower Abundance and Impaired Function of CD71+ Erythroid Cells in Inflammatory Bowel Disease Patients During Pregnancy"

    Article Title: Lower Abundance and Impaired Function of CD71+ Erythroid Cells in Inflammatory Bowel Disease Patients During Pregnancy

    Journal: Journal of Crohn's & Colitis

    doi: 10.1093/ecco-jcc/jjy147

    Expansion of CD71 + erythroid cells in human pregnancy. [A] Representative dot plots showing gating strategy and frequency of CD71 + erythroid cells in blood of pregnant versus non-pregnant women. [B] Cumulative data indicating percentages of CD71 + erythroid cells in preconception [PC], first trimester [T1], second trimester [T2], third trimester [T3], and postpartum [PP]. [C] Representative dot plots showing frequency of CD71 + erythroid cells in placenta tissues of a healthy woman. [D] Percentages of CD71 + erythroid cells in placental tissues of healthy women. [E] Representative dot plots showing proliferation of CD3 + T cells as measured by carboxyfluorescein succinimidyl ester [ CFSE] dye in peripheral blood mononuclear cells [PBMCs] in the presence or absence of CD71 + erythroid cells. [F] Percentage of CD3 + T cells proliferation in PBMCs in the presence or absence of CD71 + erythroid cells. [G] Representative dot plots showing IL-6 and TNF-α production among CD11b + cells in PBMCs in the presence or absence of CD71 + erythroid cells. [H] Percentages of CD11b + cells producing IL-6, TNF-α or both cytokines, in the presence or absence of CD71 + erythroid cells. [I] Representative dot plots showing proliferation of CD4 + and CD8 + T cells in the presence or absence of CD71 + erythroid cells. [J] Percentages of proliferated CD4 + and CD8 + T cells in placenta cells in the presence or absence of CD71 + erythroid cells.
    Figure Legend Snippet: Expansion of CD71 + erythroid cells in human pregnancy. [A] Representative dot plots showing gating strategy and frequency of CD71 + erythroid cells in blood of pregnant versus non-pregnant women. [B] Cumulative data indicating percentages of CD71 + erythroid cells in preconception [PC], first trimester [T1], second trimester [T2], third trimester [T3], and postpartum [PP]. [C] Representative dot plots showing frequency of CD71 + erythroid cells in placenta tissues of a healthy woman. [D] Percentages of CD71 + erythroid cells in placental tissues of healthy women. [E] Representative dot plots showing proliferation of CD3 + T cells as measured by carboxyfluorescein succinimidyl ester [ CFSE] dye in peripheral blood mononuclear cells [PBMCs] in the presence or absence of CD71 + erythroid cells. [F] Percentage of CD3 + T cells proliferation in PBMCs in the presence or absence of CD71 + erythroid cells. [G] Representative dot plots showing IL-6 and TNF-α production among CD11b + cells in PBMCs in the presence or absence of CD71 + erythroid cells. [H] Percentages of CD11b + cells producing IL-6, TNF-α or both cytokines, in the presence or absence of CD71 + erythroid cells. [I] Representative dot plots showing proliferation of CD4 + and CD8 + T cells in the presence or absence of CD71 + erythroid cells. [J] Percentages of proliferated CD4 + and CD8 + T cells in placenta cells in the presence or absence of CD71 + erythroid cells.

    Techniques Used:

    Impaired functionality of CD71 + erythroid cells from cord blood of newborns to inflammatory bowel disease [IBD] mothers. [A] Representative dot plots showing frequency of CD71 + erythroid cells in cord blood mononuclear cells [CBMCs] of infants born to ulcerative colitis [UC], Crohn’s disease [CD], and healthy control [HC] mothers. [B] Percentages of CD71 + erythroid cells in CBMCs of infants born to UC, CD, and HC mothers. [C] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to an HC mother. [D] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to HC mothers following stimulation with anti-CD3 in vitro . [E] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to a UC mother. [F] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to UC mothers. [G] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to a CD mother. [H] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to CD mothers. [I] TNF-α and [J] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to HC mothers in the presence or absence of CD71 + erythroid cells. [K] TNF-α and [L] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to UC mothers. [M] TNF-α and [N] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to CD mothers in the presence or absence of CD71 + erythroid cells as measured by ELISA.
    Figure Legend Snippet: Impaired functionality of CD71 + erythroid cells from cord blood of newborns to inflammatory bowel disease [IBD] mothers. [A] Representative dot plots showing frequency of CD71 + erythroid cells in cord blood mononuclear cells [CBMCs] of infants born to ulcerative colitis [UC], Crohn’s disease [CD], and healthy control [HC] mothers. [B] Percentages of CD71 + erythroid cells in CBMCs of infants born to UC, CD, and HC mothers. [C] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to an HC mother. [D] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to HC mothers following stimulation with anti-CD3 in vitro . [E] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to a UC mother. [F] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to UC mothers. [G] Representative dot plots showing proliferation of CD4 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of an infant born to a CD mother. [H] Percentages of proliferated CD4 + T cells and CD8 + T cells in the presence or absence of CD71 + erythroid cells among CBMCs of infants born to CD mothers. [I] TNF-α and [J] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to HC mothers in the presence or absence of CD71 + erythroid cells. [K] TNF-α and [L] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to UC mothers. [M] TNF-α and [N] IL-6 production by stimulated CBMCs with anti-CD3 antibody obtained from infants born to CD mothers in the presence or absence of CD71 + erythroid cells as measured by ELISA.

    Techniques Used: In Vitro, Enzyme-linked Immunosorbent Assay

    CD71 + erythroid cells may impact on gut haemostasis during pregnancy. [A] Representative dot plots showing percentages of CD71 + erythroid cells in placenta and [B] spleen of mice before and after anti-CD71 antibody treatment. [C] Expression of TLR-1, [D] TLR-4, [E] TLR-9, [F] TLR-2, [G] CXCL-1, [H] IL-6 and [I] TGF-β genes by gut tissues of non-pregnant compared with pregnant mice treated with rat IgG isotype control or anti-CD71 antibody three days after treatment. [J] Representa tive plots and [K] cumulative data showing IL-6 and TNF-α production by intestinal CDllc + cells. [L] Representative plots and [M] cumulative data showing IL-6 and TNF-α production by intestinal CDllb + cells. [N] Expression of VEGFα gene in placenta tissues. [O] Treated mice were subjected to 16S rRNA-based polymerase chain reaction [PCR] for total bacteria, [P] Bacteroides-Prevotella-Porphyromonas group [Q] Enterobacteriaceae group, [R] Clostridium cluster XIVa, [S] Clostridium cluster 1, and [T] Clostridium cluster IV. [U] Levels of fluorescein isothiocyanate labelled dextran [FITC-dextran] in the blood of IgG versus anti-CD71 treated mice. Data are obtained from a minimum of five mice/group and at least two independent experiments.
    Figure Legend Snippet: CD71 + erythroid cells may impact on gut haemostasis during pregnancy. [A] Representative dot plots showing percentages of CD71 + erythroid cells in placenta and [B] spleen of mice before and after anti-CD71 antibody treatment. [C] Expression of TLR-1, [D] TLR-4, [E] TLR-9, [F] TLR-2, [G] CXCL-1, [H] IL-6 and [I] TGF-β genes by gut tissues of non-pregnant compared with pregnant mice treated with rat IgG isotype control or anti-CD71 antibody three days after treatment. [J] Representa tive plots and [K] cumulative data showing IL-6 and TNF-α production by intestinal CDllc + cells. [L] Representative plots and [M] cumulative data showing IL-6 and TNF-α production by intestinal CDllb + cells. [N] Expression of VEGFα gene in placenta tissues. [O] Treated mice were subjected to 16S rRNA-based polymerase chain reaction [PCR] for total bacteria, [P] Bacteroides-Prevotella-Porphyromonas group [Q] Enterobacteriaceae group, [R] Clostridium cluster XIVa, [S] Clostridium cluster 1, and [T] Clostridium cluster IV. [U] Levels of fluorescein isothiocyanate labelled dextran [FITC-dextran] in the blood of IgG versus anti-CD71 treated mice. Data are obtained from a minimum of five mice/group and at least two independent experiments.

    Techniques Used: Mouse Assay, Expressing, Polymerase Chain Reaction

    24) Product Images from "MicroRNA-140 mediates RB tumor suppressor function to control stem cell-like activity through interleukin-6"

    Article Title: MicroRNA-140 mediates RB tumor suppressor function to control stem cell-like activity through interleukin-6

    Journal: Oncotarget

    doi: 10.18632/oncotarget.14681

    mmu-mir-140 mediates Rb function to control Il-6 expression ( A ) RT-qPCR of Il-6 in p 53-null soft tissue sarcoma cells transduced with the indicated shRNA and retroviral vectors. N = 3. ( B ) RT-qPCR of Il-6 in p 53-null soft tissue sarcoma cells transduced with the indicated vectors. N = 3. ( C ) The sequence of the mouse Il-6 3′UTR containing seed sequence of mmu-mir-140 (underlined). The seed sequence of the mouse Il-6 3′UTR was mutated as shown (mIl-6-3′ UTR Mut ). ( D ) m Il-6-3Times New RomanUTR WT or m Il-6-3′UTR Mut was transduced into NIH3T3 cells together with the mmu-mir-140 expression vector or control (scramble). After 48 hours, luciferase activity was measured. ( E ) Number of spheres derived from 5 × 10 4 of p 53-null soft tissue sarcoma cells transduced with the indicated vector in the presence of indicated concentration of recombinant mouse Il-6 (#406-ML-005, R D Systems). N = 3.
    Figure Legend Snippet: mmu-mir-140 mediates Rb function to control Il-6 expression ( A ) RT-qPCR of Il-6 in p 53-null soft tissue sarcoma cells transduced with the indicated shRNA and retroviral vectors. N = 3. ( B ) RT-qPCR of Il-6 in p 53-null soft tissue sarcoma cells transduced with the indicated vectors. N = 3. ( C ) The sequence of the mouse Il-6 3′UTR containing seed sequence of mmu-mir-140 (underlined). The seed sequence of the mouse Il-6 3′UTR was mutated as shown (mIl-6-3′ UTR Mut ). ( D ) m Il-6-3Times New RomanUTR WT or m Il-6-3′UTR Mut was transduced into NIH3T3 cells together with the mmu-mir-140 expression vector or control (scramble). After 48 hours, luciferase activity was measured. ( E ) Number of spheres derived from 5 × 10 4 of p 53-null soft tissue sarcoma cells transduced with the indicated vector in the presence of indicated concentration of recombinant mouse Il-6 (#406-ML-005, R D Systems). N = 3.

    Techniques Used: Expressing, Quantitative RT-PCR, Transduction, shRNA, Sequencing, Plasmid Preparation, Luciferase, Activity Assay, Derivative Assay, Concentration Assay, Recombinant

    Identification of genes induced by Rb depletion in a mir-140 -dependent manner ( A ) The ontology of the top 3 gene sets influenced by Rb depletion possibly in an mir -140-dependent manner was determined by GO Biological Process provided by DAVID. ( B ) A heat map illustrating the expression levels of the indicated genes in p 53-null soft tissue sarcoma cells transduced with the indicated vector. ( C ) The sequences of mouse Il-6, Vegfa, Wnt5a and Serping1 3′UTR corresponding to the seed sequence of mmu-mir-140 (underline) are presented with their mirSVR and PhastCons scores.
    Figure Legend Snippet: Identification of genes induced by Rb depletion in a mir-140 -dependent manner ( A ) The ontology of the top 3 gene sets influenced by Rb depletion possibly in an mir -140-dependent manner was determined by GO Biological Process provided by DAVID. ( B ) A heat map illustrating the expression levels of the indicated genes in p 53-null soft tissue sarcoma cells transduced with the indicated vector. ( C ) The sequences of mouse Il-6, Vegfa, Wnt5a and Serping1 3′UTR corresponding to the seed sequence of mmu-mir-140 (underline) are presented with their mirSVR and PhastCons scores.

    Techniques Used: Expressing, Transduction, Plasmid Preparation, Sequencing

    hsa-mir-140 suppresses IL-6 expression induced by RB depletion in human breast cancer cells ( A ) The sequence of the human Il-6 3′UTR and the seed sequence of hsa-mir-140 (underlined) are indicated. mirSVR and Phantcons scores are indicated. ( B ) RT-qPCR of the indicated genes in MCF7 cells transduced with the indicated vector. N = 3. ( C ) IB of the indicated proteins in MCF-7 cells transduced with the indicated vector. ( D ) ELISA of human IL-6 levels in MCF-7 cells transduced with the indicated vector following 8 hours culture. ( E ) Number of spheres derived from 1.25 × 10 3 of MCF-7 cells transduced with the indicated vectors. N = 3. ( F ) Number of spheres derived from 1.25 × 10 3 of MCF-7 cells transduced with the indicated vector in the presence of indicated concentration of recombinant human IL-6 (#206-IL-010, R D Systems). N = 3. ( G ) RT-qPCR of RB7LP in MCF-7 cells transduced with the indicated vector. N = 3. ( H ) RT-qPCR of hsa-miR-140 in MCF-7 cells transduced with the indicated vector. N = 3. ( I ) h IL-6-3′UTR WT or h IL-6-3′UTR Mut was transduced into MCF-7 cells together with the hsa-mir-140 expression vector or control (scramble). After 48 hours, luciferase activity was measured.
    Figure Legend Snippet: hsa-mir-140 suppresses IL-6 expression induced by RB depletion in human breast cancer cells ( A ) The sequence of the human Il-6 3′UTR and the seed sequence of hsa-mir-140 (underlined) are indicated. mirSVR and Phantcons scores are indicated. ( B ) RT-qPCR of the indicated genes in MCF7 cells transduced with the indicated vector. N = 3. ( C ) IB of the indicated proteins in MCF-7 cells transduced with the indicated vector. ( D ) ELISA of human IL-6 levels in MCF-7 cells transduced with the indicated vector following 8 hours culture. ( E ) Number of spheres derived from 1.25 × 10 3 of MCF-7 cells transduced with the indicated vectors. N = 3. ( F ) Number of spheres derived from 1.25 × 10 3 of MCF-7 cells transduced with the indicated vector in the presence of indicated concentration of recombinant human IL-6 (#206-IL-010, R D Systems). N = 3. ( G ) RT-qPCR of RB7LP in MCF-7 cells transduced with the indicated vector. N = 3. ( H ) RT-qPCR of hsa-miR-140 in MCF-7 cells transduced with the indicated vector. N = 3. ( I ) h IL-6-3′UTR WT or h IL-6-3′UTR Mut was transduced into MCF-7 cells together with the hsa-mir-140 expression vector or control (scramble). After 48 hours, luciferase activity was measured.

    Techniques Used: Expressing, Sequencing, Quantitative RT-PCR, Transduction, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Derivative Assay, Concentration Assay, Recombinant, Luciferase, Activity Assay

    25) Product Images from "Calcineurin inhibitors cyclosporine A and tacrolimus induce vascular inflammation and endothelial activation through TLR4 signaling"

    Article Title: Calcineurin inhibitors cyclosporine A and tacrolimus induce vascular inflammation and endothelial activation through TLR4 signaling

    Journal: Scientific Reports

    doi: 10.1038/srep27915

    Pharmacological inhibition of TLR4 blocked proinflammatory responses induced by CsA and tacrolimus in endothelial cells. Cells were incubated with 10 μg/ml CsA or 20 μg/ml Tac alone or in the presence of the TLR4 inhibitor, CLI095 (added 6 h before the CNIs). ( A) Activation of NF-κB was assessed through the nuclear translocation of the NF-κB/p65 subunit detected by immunofluorescence confocal microscopy. Control cells show a cytoplasmic NF-κB/p65 staining (green) whereas in cells stimulated with CsA or Tac, NF-κB/p65 was mostly located inside nuclei. By contrast, the TLR4 inhibitor CLI-095 prevented nuclear translocation of NF-κB/p65. Cells were also stimulated with TNF-α (30 ng/ml) alone as a positive control of NF-κB activation or preincubated with CLI-095 to prove that TLR4 inhibition does not interfere with TNF-α-mediated NF-κB/p65 translocation. ( B,C) Transcriptional levels of proinflammatory cytokines (CCL2, CCL5, TNF-α, IL-6) ( A ) and endothelial activation markers (ICAM-1, VCAM1, SELE) ( B ) were evaluated by qRT-PCR. Data are mean ± SEM of three independent experiments. *p ≤ 0.05 vs control; # p ≤ 0.05 and † p ≤ 0.05 vs CsA or Tac, respectively.
    Figure Legend Snippet: Pharmacological inhibition of TLR4 blocked proinflammatory responses induced by CsA and tacrolimus in endothelial cells. Cells were incubated with 10 μg/ml CsA or 20 μg/ml Tac alone or in the presence of the TLR4 inhibitor, CLI095 (added 6 h before the CNIs). ( A) Activation of NF-κB was assessed through the nuclear translocation of the NF-κB/p65 subunit detected by immunofluorescence confocal microscopy. Control cells show a cytoplasmic NF-κB/p65 staining (green) whereas in cells stimulated with CsA or Tac, NF-κB/p65 was mostly located inside nuclei. By contrast, the TLR4 inhibitor CLI-095 prevented nuclear translocation of NF-κB/p65. Cells were also stimulated with TNF-α (30 ng/ml) alone as a positive control of NF-κB activation or preincubated with CLI-095 to prove that TLR4 inhibition does not interfere with TNF-α-mediated NF-κB/p65 translocation. ( B,C) Transcriptional levels of proinflammatory cytokines (CCL2, CCL5, TNF-α, IL-6) ( A ) and endothelial activation markers (ICAM-1, VCAM1, SELE) ( B ) were evaluated by qRT-PCR. Data are mean ± SEM of three independent experiments. *p ≤ 0.05 vs control; # p ≤ 0.05 and † p ≤ 0.05 vs CsA or Tac, respectively.

    Techniques Used: Inhibition, Incubation, Activation Assay, Translocation Assay, Immunofluorescence, Confocal Microscopy, Staining, Positive Control, Quantitative RT-PCR

    CNIs induce the expression of inflammatory mediators and endothelial activation markers in murine endothelial cells. The expression of proinflammatory cytokines and endothelial activation markers was evaluated in murine endothelial cells exposed to cyclosporine (CsA) or tacrolimus (Tac). ( A) Dose-dependent CCL2 and CCL5 mRNA expression in cells treated for 6 h with 10–20 μg/ml CsA or Tac, assessed by qRT-PCR. ( B) TNF-α, IL-6, ICAM-1 and VCAM-1 mRNA expression in cells treated with 10 μg/ml CsA and 20 μg/ml Tac, qRT-PCR. ( C,D) CCL2 ( C ) or ICAM-1 ( D ) protein levels assessed by ELISA in supernatants from cells stimulated with 10 μg/ml CsA, 20 μg/ml Tac or vehicle (Control). Data represent the mean ± SEM of three independent experiments. *p ≤ 0.05 vs control.
    Figure Legend Snippet: CNIs induce the expression of inflammatory mediators and endothelial activation markers in murine endothelial cells. The expression of proinflammatory cytokines and endothelial activation markers was evaluated in murine endothelial cells exposed to cyclosporine (CsA) or tacrolimus (Tac). ( A) Dose-dependent CCL2 and CCL5 mRNA expression in cells treated for 6 h with 10–20 μg/ml CsA or Tac, assessed by qRT-PCR. ( B) TNF-α, IL-6, ICAM-1 and VCAM-1 mRNA expression in cells treated with 10 μg/ml CsA and 20 μg/ml Tac, qRT-PCR. ( C,D) CCL2 ( C ) or ICAM-1 ( D ) protein levels assessed by ELISA in supernatants from cells stimulated with 10 μg/ml CsA, 20 μg/ml Tac or vehicle (Control). Data represent the mean ± SEM of three independent experiments. *p ≤ 0.05 vs control.

    Techniques Used: Expressing, Activation Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    CNI induce inflammation in wild-type aortas but not in aortas from TLR4 −/− mice. Aorta tissue segments extracted from wild-type or TLR4 −/− C57BL/6 mice were stimulated for 6 h with 10 μg/ml CsA or 20 μg/ml Tac. CLI-095 was added 6 h before stimulation with the CNIs. ( A,B) Confocal microphotographs showing NF-κB/p65 content and location in control or CsA or CsA plus CLI-095 treated aortic sections from wild-type mice. Activation of NF-κB/p65 was detected by intensification of the specific red fluorescence in cytoplasm and nucleus of either endothelial cells recognized by CD31 staining (green fluorescence surrounding the cell borders) ( A ) or VSMC cells expressing αSMA (cytoplasmic green fluorescence) ( B ). Endothelial cells were found lining the intima layer and facing the lumen and VSMC located in the media layer. Yellow asterisks point cells with nuclear translocation of p65 and white asterisks indicate cells with increased cytoplasmic p65 expression. White arrows point endothelial and VSMC without increased expression of p65 in control or CLI095 treated aortas. White arrowheads in A show elastin fibers (green autofluorescence) which otherwise are not apparently visualized in B because the much higher αSMA specific fluorescence. Nuclei were counterstained with DAPI. Original magnification x630. ( C,D) Gene expression of CCL2, CCL5, IL-6, TNF-α (left panel) and ICAM-1, ET-1(right panel) in cultured aorta sections from wild-type mice exposed to CsA or Tac alone or in the presence of CLI-095. Data are expressed as mean ± SEM of 4 samples. *p
    Figure Legend Snippet: CNI induce inflammation in wild-type aortas but not in aortas from TLR4 −/− mice. Aorta tissue segments extracted from wild-type or TLR4 −/− C57BL/6 mice were stimulated for 6 h with 10 μg/ml CsA or 20 μg/ml Tac. CLI-095 was added 6 h before stimulation with the CNIs. ( A,B) Confocal microphotographs showing NF-κB/p65 content and location in control or CsA or CsA plus CLI-095 treated aortic sections from wild-type mice. Activation of NF-κB/p65 was detected by intensification of the specific red fluorescence in cytoplasm and nucleus of either endothelial cells recognized by CD31 staining (green fluorescence surrounding the cell borders) ( A ) or VSMC cells expressing αSMA (cytoplasmic green fluorescence) ( B ). Endothelial cells were found lining the intima layer and facing the lumen and VSMC located in the media layer. Yellow asterisks point cells with nuclear translocation of p65 and white asterisks indicate cells with increased cytoplasmic p65 expression. White arrows point endothelial and VSMC without increased expression of p65 in control or CLI095 treated aortas. White arrowheads in A show elastin fibers (green autofluorescence) which otherwise are not apparently visualized in B because the much higher αSMA specific fluorescence. Nuclei were counterstained with DAPI. Original magnification x630. ( C,D) Gene expression of CCL2, CCL5, IL-6, TNF-α (left panel) and ICAM-1, ET-1(right panel) in cultured aorta sections from wild-type mice exposed to CsA or Tac alone or in the presence of CLI-095. Data are expressed as mean ± SEM of 4 samples. *p

    Techniques Used: Mouse Assay, Activation Assay, Fluorescence, Staining, Expressing, Translocation Assay, Cell Culture

    26) Product Images from "Differential modulation of Ahr and Arid5a: A promising therapeutic strategy for autoimmune encephalomyelitis"

    Article Title: Differential modulation of Ahr and Arid5a: A promising therapeutic strategy for autoimmune encephalomyelitis

    Journal: Saudi Pharmaceutical Journal : SPJ

    doi: 10.1016/j.jsps.2020.10.007

    The modulatory effects of Flavipin on cytokines are mediated by Ahr- and Arid5a-dependent effects. Naïve T cells (CD4 + CD62L + ) were cultured under Th17- or Treg-inducing conditions, and peritoneal macrophages were stimulated with LPS in presence or absence of Flavipin (40 μM). The cells were electroporated with nonspecific siRNA (siNS; 100 nM) or Ahr siRNA (siAhr; 100 nM), siArid5a or. Cytokines were measured using ELISA, and mRNAs were measured by real‐time PCR and normalized to Gapdh mRNA. (A) Levels of IL‐17 and TGF‐β produced by CD4 + T cells cultured under Th17- or Treg-inducing conditions (72 h), respectively. (B) Levels of IL-6, TNF-α and IL-23 produced by LPS-stimulated macrophages (16 h). (C and E) Levels of IL‐17 and TGF‐β produced by CD4 + T cells electroporated with siAhr, siArid5a or siNS and cultured under Th17- or Treg-inducing conditions. (D and F) Levels of IL-6, TNF-α and IL-23 produced by macrophages electroporated with siAhr, siArid5a or siNS and stimulated with LPS. (G) Relative mRNA expression of Stat3 and OX40 in CD4 + T cells cultured under Th17-inducing conditions (12 h) compared with DMSO-treated cells. (H) The efficiency of siArid5a was confirmed by immunoblot. (I) Clinical and maximum scores of EAE induced in Rag‐1 −/− mice by adoptive transfer of encephalitogenic CD4 + T cells isolated from EAE mice (10 days post immunization) and restimulated with MOG 35–55 (72 h). Data in A-G are shown as the mean ± SD of representative experiment out of three independent experiments ( n = 3 each) studied in triplicates and produced similar results. Data in I (left panel) are shown as the mean ± SD from representative experiment out of two independent experiments (n = 5 each) produced similar results; I (right panel) represents individual animals. * p
    Figure Legend Snippet: The modulatory effects of Flavipin on cytokines are mediated by Ahr- and Arid5a-dependent effects. Naïve T cells (CD4 + CD62L + ) were cultured under Th17- or Treg-inducing conditions, and peritoneal macrophages were stimulated with LPS in presence or absence of Flavipin (40 μM). The cells were electroporated with nonspecific siRNA (siNS; 100 nM) or Ahr siRNA (siAhr; 100 nM), siArid5a or. Cytokines were measured using ELISA, and mRNAs were measured by real‐time PCR and normalized to Gapdh mRNA. (A) Levels of IL‐17 and TGF‐β produced by CD4 + T cells cultured under Th17- or Treg-inducing conditions (72 h), respectively. (B) Levels of IL-6, TNF-α and IL-23 produced by LPS-stimulated macrophages (16 h). (C and E) Levels of IL‐17 and TGF‐β produced by CD4 + T cells electroporated with siAhr, siArid5a or siNS and cultured under Th17- or Treg-inducing conditions. (D and F) Levels of IL-6, TNF-α and IL-23 produced by macrophages electroporated with siAhr, siArid5a or siNS and stimulated with LPS. (G) Relative mRNA expression of Stat3 and OX40 in CD4 + T cells cultured under Th17-inducing conditions (12 h) compared with DMSO-treated cells. (H) The efficiency of siArid5a was confirmed by immunoblot. (I) Clinical and maximum scores of EAE induced in Rag‐1 −/− mice by adoptive transfer of encephalitogenic CD4 + T cells isolated from EAE mice (10 days post immunization) and restimulated with MOG 35–55 (72 h). Data in A-G are shown as the mean ± SD of representative experiment out of three independent experiments ( n = 3 each) studied in triplicates and produced similar results. Data in I (left panel) are shown as the mean ± SD from representative experiment out of two independent experiments (n = 5 each) produced similar results; I (right panel) represents individual animals. * p

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Produced, Expressing, Mouse Assay, Adoptive Transfer Assay, Isolation

    Arid5a stabilizing functions and competition with Regnase-1 on the 3′UTR of target mRNAs are abolished by Flavipin . (A) Relative luciferase activity of pGL3 plasmids (100 ng) encoding the 3′UTR of Il6 , Stat3 or OX40 (1076‐1119) transfected into HEK293T cells (48 h) together with Arid5a expression plasmid (300 ng) compared with each activity when transfected with empty plasmid (Control; 300 ng); Flavipin (40 μM). (B) Relative luciferase activity of pGL3 plasmids (100 ng) encoding the 3′UTR of Il6 , Stat3 and OX40 (1076‐1119) transfected into HEK293T cells (48 h) together with Arid5a (300 ng) and/or Regnase-1 (300 ng) expression plasmids compared with each activity when transfected with empty plasmid (Control; 300–600 ng); Flavipin (40 μM). Data are shown as the mean ± SD from representative experiment out of three independent experiments studied in triplicates and produced similar results. * p
    Figure Legend Snippet: Arid5a stabilizing functions and competition with Regnase-1 on the 3′UTR of target mRNAs are abolished by Flavipin . (A) Relative luciferase activity of pGL3 plasmids (100 ng) encoding the 3′UTR of Il6 , Stat3 or OX40 (1076‐1119) transfected into HEK293T cells (48 h) together with Arid5a expression plasmid (300 ng) compared with each activity when transfected with empty plasmid (Control; 300 ng); Flavipin (40 μM). (B) Relative luciferase activity of pGL3 plasmids (100 ng) encoding the 3′UTR of Il6 , Stat3 and OX40 (1076‐1119) transfected into HEK293T cells (48 h) together with Arid5a (300 ng) and/or Regnase-1 (300 ng) expression plasmids compared with each activity when transfected with empty plasmid (Control; 300–600 ng); Flavipin (40 μM). Data are shown as the mean ± SD from representative experiment out of three independent experiments studied in triplicates and produced similar results. * p

    Techniques Used: Luciferase, Activity Assay, Transfection, Expressing, Plasmid Preparation, Produced

    Ahr- and Arid5a-dependent actions mediate the alleviating effects of Flavipin on EAE . MOG 35‐55 emulsified in CFA and pertussis toxin were used to induce EAE. The cytokines were quantified by ELISA, and the expression of mRNA encoding protein and miRNAs was assessed by real‐time PCR and normalized to Gapdh or RNU6B mRNAs, respectively. (A) EAE clinical and maximum score and (B) incidence (%) in vehicle (Control)- or Flavipin (Flav)-treated mice. (C) Levels of IL‐6 and TNF‐α in the serum (day 23 post immunization). (D) Levels of IL‐17 and TGF‐β produced by encephalitogenic CD4 + T cells isolated from the inguinal lymph nodes (10 days post immunization) and restimulated with MOG 35–55 (72 h). (E) Flow cytometry analysis of the frequency (%) of CD4 + IL‐17 + and CD4 + FoxP3 + T cells in the inguinal lymph nodes (day 23 post immunization) and the frequency of CD4 + OX40 + T cells in the CNS (10 days post immunization), gated on CD4 + T cells. (F) Absolute number of CD4 + CD45 + T cells in the CNS (day 10 post immunization). (G) Culture supernatant level of IL‐17 produced by encephalitogenic CD4 + T cells restimulated as described in D in the absence (Control) or presence of agonistic anti-OX40 antibodies (OX86). (H) Relative expression of Cyp1A1 mRNA in CD4 + T cells and CD11b + macrophages isolated from the spleens of EAE mice (day 10 post immunization) compared to the control EAE mice. (I) Relative expression of miR-132 and miR-212 in CD4 + T cells and CD11b + macrophages isolated from the spleens of EAE mice (10 post immunization) compared to the control EAE mice. (J) Relative expression of AChE mRNA in splenocytes from EAE mice (day 10 post immunization) compared to control EAE mice. Data in A and C-J are shown as the mean ± SD from representative experiment out of three independent experiments ( n = 5 each) produced similar results; C-J were studied in triplicates. * p
    Figure Legend Snippet: Ahr- and Arid5a-dependent actions mediate the alleviating effects of Flavipin on EAE . MOG 35‐55 emulsified in CFA and pertussis toxin were used to induce EAE. The cytokines were quantified by ELISA, and the expression of mRNA encoding protein and miRNAs was assessed by real‐time PCR and normalized to Gapdh or RNU6B mRNAs, respectively. (A) EAE clinical and maximum score and (B) incidence (%) in vehicle (Control)- or Flavipin (Flav)-treated mice. (C) Levels of IL‐6 and TNF‐α in the serum (day 23 post immunization). (D) Levels of IL‐17 and TGF‐β produced by encephalitogenic CD4 + T cells isolated from the inguinal lymph nodes (10 days post immunization) and restimulated with MOG 35–55 (72 h). (E) Flow cytometry analysis of the frequency (%) of CD4 + IL‐17 + and CD4 + FoxP3 + T cells in the inguinal lymph nodes (day 23 post immunization) and the frequency of CD4 + OX40 + T cells in the CNS (10 days post immunization), gated on CD4 + T cells. (F) Absolute number of CD4 + CD45 + T cells in the CNS (day 10 post immunization). (G) Culture supernatant level of IL‐17 produced by encephalitogenic CD4 + T cells restimulated as described in D in the absence (Control) or presence of agonistic anti-OX40 antibodies (OX86). (H) Relative expression of Cyp1A1 mRNA in CD4 + T cells and CD11b + macrophages isolated from the spleens of EAE mice (day 10 post immunization) compared to the control EAE mice. (I) Relative expression of miR-132 and miR-212 in CD4 + T cells and CD11b + macrophages isolated from the spleens of EAE mice (10 post immunization) compared to the control EAE mice. (J) Relative expression of AChE mRNA in splenocytes from EAE mice (day 10 post immunization) compared to control EAE mice. Data in A and C-J are shown as the mean ± SD from representative experiment out of three independent experiments ( n = 5 each) produced similar results; C-J were studied in triplicates. * p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Produced, Isolation, Flow Cytometry

    27) Product Images from "Sildenafil ameliorates right ventricular early molecular derangement during left ventricular pressure overload"

    Article Title: Sildenafil ameliorates right ventricular early molecular derangement during left ventricular pressure overload

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0195528

    Pathological gene induction in RV as well as LV were ameliorated by sildenafil. (A) Expression of fetal genes as markers of cardiac hypertrophy, encoding for BNP and B-MHC, normalized to GAPDH. Transverse aortic constriction (TAC) for two days induced marked increase in BNP mRNA levels in the RV myocardium as well as in the LV myocardium, which was prevented by sildenafil. Expression of B-MHC was increased by two-day TAC in LV, but was not affected by sildenafil. (B) Expression of inflammatory cytokines, IL1b and IL6, normalized to GAPDH. IL1b and IL6 was up-regulated by TAC in both ventricles. Sildenafil inhibited both in LV, and also attenuated both in RV. (C) Expression of genes for enzymes inducing oxidative stress, NOX2 and NOX4, normalized to GAPDH. Expression of NOX2 was increased by TAC in both ventricles, and suppressed by sildenafil in LV. TAC and sildenafil had little influence on NOX4 expression. Results are expressed as mean ± s.e.m. (n = 5). TAC 2d Veh, TAC for 2 days with vehicle treatment; TAC 2d Sil, TAC for 2days with sildenafil treatment. n.s., not significant by one-way analysis of variance; *, p
    Figure Legend Snippet: Pathological gene induction in RV as well as LV were ameliorated by sildenafil. (A) Expression of fetal genes as markers of cardiac hypertrophy, encoding for BNP and B-MHC, normalized to GAPDH. Transverse aortic constriction (TAC) for two days induced marked increase in BNP mRNA levels in the RV myocardium as well as in the LV myocardium, which was prevented by sildenafil. Expression of B-MHC was increased by two-day TAC in LV, but was not affected by sildenafil. (B) Expression of inflammatory cytokines, IL1b and IL6, normalized to GAPDH. IL1b and IL6 was up-regulated by TAC in both ventricles. Sildenafil inhibited both in LV, and also attenuated both in RV. (C) Expression of genes for enzymes inducing oxidative stress, NOX2 and NOX4, normalized to GAPDH. Expression of NOX2 was increased by TAC in both ventricles, and suppressed by sildenafil in LV. TAC and sildenafil had little influence on NOX4 expression. Results are expressed as mean ± s.e.m. (n = 5). TAC 2d Veh, TAC for 2 days with vehicle treatment; TAC 2d Sil, TAC for 2days with sildenafil treatment. n.s., not significant by one-way analysis of variance; *, p

    Techniques Used: Expressing

    Dexamethasone suppressed pathological molecular remodeling induced in the RV as well as the LV. (A) mRNA expression of IL1b and IL6, normalized to GAPDH. Dexamethasone inhibited overexpression of IL1b in both RV and LV myocardium induced by transverse aortic constriction (TAC) for 2 days. (B) mRNA expression of RCAN1 and BNP normalized to GAPDH. Dexamethasone suppressed up-regulation of RCAN1 and BNP in RV and LV myocardium induced by two-day TAC. Results are expressed as mean ± s.e.m. (n = 5). TAC 2d Veh, TAC for 2 days with vehicle treatment; TAC 2d DXM, TAC for 2 days with dexamethasone treatment. n.s. *, p
    Figure Legend Snippet: Dexamethasone suppressed pathological molecular remodeling induced in the RV as well as the LV. (A) mRNA expression of IL1b and IL6, normalized to GAPDH. Dexamethasone inhibited overexpression of IL1b in both RV and LV myocardium induced by transverse aortic constriction (TAC) for 2 days. (B) mRNA expression of RCAN1 and BNP normalized to GAPDH. Dexamethasone suppressed up-regulation of RCAN1 and BNP in RV and LV myocardium induced by two-day TAC. Results are expressed as mean ± s.e.m. (n = 5). TAC 2d Veh, TAC for 2 days with vehicle treatment; TAC 2d DXM, TAC for 2 days with dexamethasone treatment. n.s. *, p

    Techniques Used: Expressing, Over Expression

    28) Product Images from "Dietary fats promote functional and structural changes in the median eminence blood/spinal fluid interface—the protective role for BDNF"

    Article Title: Dietary fats promote functional and structural changes in the median eminence blood/spinal fluid interface—the protective role for BDNF

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-1046-8

    Modulating BDNF in obese-prone and obese-resistant mice. Schematic representation of the protocol employed to modulate BDNF in obese-prone (OP) and obese-resistant (OR) mice ( a ). Body mass variation ( b ), percent fat mass as determined by PET-CT scan ( c ), and mean daily food intake ( d ) in the experimental models. In e – h , real-time PCR determination of the expression of transcripts encoding for IGFBP2 ( e ), NGFR ( f ), TNFα ( g ), and IL6 ( h ) in the median eminence of mice. In e – h , n = 4; * p
    Figure Legend Snippet: Modulating BDNF in obese-prone and obese-resistant mice. Schematic representation of the protocol employed to modulate BDNF in obese-prone (OP) and obese-resistant (OR) mice ( a ). Body mass variation ( b ), percent fat mass as determined by PET-CT scan ( c ), and mean daily food intake ( d ) in the experimental models. In e – h , real-time PCR determination of the expression of transcripts encoding for IGFBP2 ( e ), NGFR ( f ), TNFα ( g ), and IL6 ( h ) in the median eminence of mice. In e – h , n = 4; * p

    Techniques Used: Mouse Assay, Positron Emission Tomography, Computed Tomography, Real-time Polymerase Chain Reaction, Expressing

    Immunoneutralization of BDNF. Schematic representation of the protocol employed to immunoneutralize BDNF ( a ). b Body mass during the experimental period. c Body mass variation during the experimental period. d Mean daily food intake during the experimental period. e – h Real-time PCR determination of the expression of transcripts encoding for TNFα ( e ), IL6 ( f ), IGFBP2 ( g ), and GFAP ( h ) in the median eminence of mice. i Immunofluorescence staining using primary antibodies against IGFBP2 (green) and BDNF (red). Specimens were obtained from bregma-anteroposterior − 2.06 to − 2.18. The images are representative of three independent experiments. In e – h , n = 4; * p
    Figure Legend Snippet: Immunoneutralization of BDNF. Schematic representation of the protocol employed to immunoneutralize BDNF ( a ). b Body mass during the experimental period. c Body mass variation during the experimental period. d Mean daily food intake during the experimental period. e – h Real-time PCR determination of the expression of transcripts encoding for TNFα ( e ), IL6 ( f ), IGFBP2 ( g ), and GFAP ( h ) in the median eminence of mice. i Immunofluorescence staining using primary antibodies against IGFBP2 (green) and BDNF (red). Specimens were obtained from bregma-anteroposterior − 2.06 to − 2.18. The images are representative of three independent experiments. In e – h , n = 4; * p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Mouse Assay, Immunofluorescence, Staining

    29) Product Images from "Inhibition of Allergic Response by Intranasal Selective NF-κB Decoy Oligodeoxynucleotides in a Murine Model of Allergic Rhinitis"

    Article Title: Inhibition of Allergic Response by Intranasal Selective NF-κB Decoy Oligodeoxynucleotides in a Murine Model of Allergic Rhinitis

    Journal: Allergy, Asthma & Immunology Research

    doi: 10.4168/aair.2017.9.1.61

    Expression of IL-4 (A), IL-5 (B), IL-6 (C), TNF-α (D), and IFN-γ (E) in the nasal mucosa by real time PCR. The transcriptional levels of IL-5 and TNF-α were decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). * P
    Figure Legend Snippet: Expression of IL-4 (A), IL-5 (B), IL-6 (C), TNF-α (D), and IFN-γ (E) in the nasal mucosa by real time PCR. The transcriptional levels of IL-5 and TNF-α were decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Positive Control

    Concentrations of IL-6 in nasal lavage fluid of each group by ELISA. The levels of IL-6 were significantly decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). * P
    Figure Legend Snippet: Concentrations of IL-6 in nasal lavage fluid of each group by ELISA. The levels of IL-6 were significantly decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). * P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control

    30) Product Images from "Knockout of receptor for advanced glycation end‐products attenuates age‐related renal lesions, et al. Knockout of receptor for advanced glycation end‐products attenuates age‐related renal lesions"

    Article Title: Knockout of receptor for advanced glycation end‐products attenuates age‐related renal lesions, et al. Knockout of receptor for advanced glycation end‐products attenuates age‐related renal lesions

    Journal: Aging Cell

    doi: 10.1111/acel.12850

    Inflammation and oxidation markers were decreased in RAGE −/− mice. Inflammation markers (a) Il‐6 , (b) Tnfα and (c) Vcam‐1 and oxidation markers (d) Sod2 , (e) Cat and (f) Hmox mRNA expression were measured in renal tissue from 3‐ or 20‐month‐old WT and RAGE −/− mice (mean ±SEM, n = 3 for 3‐month‐old mice and n = 10 for 20‐month‐old mice). Representative western blot and quantification of protein levels of (g) SIRT1 (mean ±SEM, n = 10), (h) pS6RP (Ser235/236)/S6RP and (i) pAKT (Ser473)/AKT in kidney extracts of 22‐ to 26‐month‐old mice (mean ±SEM, n = 4). * p
    Figure Legend Snippet: Inflammation and oxidation markers were decreased in RAGE −/− mice. Inflammation markers (a) Il‐6 , (b) Tnfα and (c) Vcam‐1 and oxidation markers (d) Sod2 , (e) Cat and (f) Hmox mRNA expression were measured in renal tissue from 3‐ or 20‐month‐old WT and RAGE −/− mice (mean ±SEM, n = 3 for 3‐month‐old mice and n = 10 for 20‐month‐old mice). Representative western blot and quantification of protein levels of (g) SIRT1 (mean ±SEM, n = 10), (h) pS6RP (Ser235/236)/S6RP and (i) pAKT (Ser473)/AKT in kidney extracts of 22‐ to 26‐month‐old mice (mean ±SEM, n = 4). * p

    Techniques Used: Mouse Assay, Expressing, Western Blot

    31) Product Images from "Immunotoxicity and allergenic potential induced by topical application of perfluorooctanoic acid (PFOA) in a murine model"

    Article Title: Immunotoxicity and allergenic potential induced by topical application of perfluorooctanoic acid (PFOA) in a murine model

    Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

    doi: 10.1016/j.fct.2020.111114

    Skin gene expression following dermal exposure to PFOA. Gene expression in the skin following 4 and 14 days of PFOA exposure. Changes in Tslp (A), Il-1beta (B), Il-6 (C), PPARα (D), Nfkb1 (E), Flg2 (F), Lor (G), and Itgbl1 (H) were evaluated. Bars represent mean (± SE) of 5 mice per group. Levels of statistical significance are denoted (**p
    Figure Legend Snippet: Skin gene expression following dermal exposure to PFOA. Gene expression in the skin following 4 and 14 days of PFOA exposure. Changes in Tslp (A), Il-1beta (B), Il-6 (C), PPARα (D), Nfkb1 (E), Flg2 (F), Lor (G), and Itgbl1 (H) were evaluated. Bars represent mean (± SE) of 5 mice per group. Levels of statistical significance are denoted (**p

    Techniques Used: Expressing, Mouse Assay

    32) Product Images from "Viral RNA and DNA Trigger Common Antiviral Responses in Mesangial Cells"

    Article Title: Viral RNA and DNA Trigger Common Antiviral Responses in Mesangial Cells

    Journal:

    doi: 10.1681/ASN.2008101067

    Dai is dispensable for non-CpG-DNA recognition in MCs. (A) Mouse MCs were transfected with Dai (Zbp1)-specific siRNA or nonspecific control siRNA and subjected real-time RT-PCR to evaluate the expression of Dai. (B) The mRNA expression of Cxcl10 and Il-6
    Figure Legend Snippet: Dai is dispensable for non-CpG-DNA recognition in MCs. (A) Mouse MCs were transfected with Dai (Zbp1)-specific siRNA or nonspecific control siRNA and subjected real-time RT-PCR to evaluate the expression of Dai. (B) The mRNA expression of Cxcl10 and Il-6

    Techniques Used: Transfection, Quantitative RT-PCR, Expressing

    3P-RNA and non-CpG-DNA complexed with CL induce Il-6 release in MCs. (A) Primary MCs were stimulated with increasing doses of 3P-RNA/CL and non-CpG-DNA/CL complexes. The highest doses also were tested in the absence of CL (−CL). PolyI:polyC RNA
    Figure Legend Snippet: 3P-RNA and non-CpG-DNA complexed with CL induce Il-6 release in MCs. (A) Primary MCs were stimulated with increasing doses of 3P-RNA/CL and non-CpG-DNA/CL complexes. The highest doses also were tested in the absence of CL (−CL). PolyI:polyC RNA

    Techniques Used:

    33) Product Images from "Autoimmunity in Dry Eye is due to Resistance of Th17 to Treg Suppression 1"

    Article Title: Autoimmunity in Dry Eye is due to Resistance of Th17 to Treg Suppression 1

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    Detection of Th17 cells and IL-17 receptor ( a ) Draining LN cells of normal and dry eye mice were stimulated with PMA+ionomycin for 6 hr and analyzed for intracellular IL-17A in CD4+ T cell population by flow cytometry (p = 0.026). ( b ) Real-time PCR analysis of the draining LN and ( c ) conjunctiva of normal and dry eye mice showing expression levels of IL-17A, IL-6 and Foxp3 mRNAs. ( d ) Representative micrograph of immunostaining of cross sections of eye showing expression of IL-17 receptor (IL-17RA) by the corneal and conjunctival epithelia of normal and dry eye mice. ( e ) Real-time PCR analysis showing IL-17RA mRNA expression levels in the cornea and conjunctiva of normal and dry eye mice.
    Figure Legend Snippet: Detection of Th17 cells and IL-17 receptor ( a ) Draining LN cells of normal and dry eye mice were stimulated with PMA+ionomycin for 6 hr and analyzed for intracellular IL-17A in CD4+ T cell population by flow cytometry (p = 0.026). ( b ) Real-time PCR analysis of the draining LN and ( c ) conjunctiva of normal and dry eye mice showing expression levels of IL-17A, IL-6 and Foxp3 mRNAs. ( d ) Representative micrograph of immunostaining of cross sections of eye showing expression of IL-17 receptor (IL-17RA) by the corneal and conjunctival epithelia of normal and dry eye mice. ( e ) Real-time PCR analysis showing IL-17RA mRNA expression levels in the cornea and conjunctiva of normal and dry eye mice.

    Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, Expressing, Immunostaining

    34) Product Images from "Contact Hypersensitivity to Oxazolone Provokes Vulvar Mechanical Hyperalgesia in Mice"

    Article Title: Contact Hypersensitivity to Oxazolone Provokes Vulvar Mechanical Hyperalgesia in Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0078673

    Labiar oxazolone challenge induces acute vulvar mechanical hyperalgesia in sensitized female ND4 Swiss mice. Mice that received a single oxazolone challenge following sensitization show increased mechanical hyperalgesia (A), increased myeloperoxidase activity in the labiar tissue (B), and increased abundance of CXCL2 , IL-6 , CXCL-1 , IFN-γ , IL-1β , and TNF-α mRNAs (C). Significances are compared to Ox/EtOH (* = p
    Figure Legend Snippet: Labiar oxazolone challenge induces acute vulvar mechanical hyperalgesia in sensitized female ND4 Swiss mice. Mice that received a single oxazolone challenge following sensitization show increased mechanical hyperalgesia (A), increased myeloperoxidase activity in the labiar tissue (B), and increased abundance of CXCL2 , IL-6 , CXCL-1 , IFN-γ , IL-1β , and TNF-α mRNAs (C). Significances are compared to Ox/EtOH (* = p

    Techniques Used: Mouse Assay, Activity Assay

    35) Product Images from "The Time of Prenatal Immune Challenge Determines the Specificity of Inflammation-Mediated Brain and Behavioral Pathology"

    Article Title: The Time of Prenatal Immune Challenge Determines the Specificity of Inflammation-Mediated Brain and Behavioral Pathology

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0099-06.2006

    Comparison of the maternal serum cytokine responses after immune challenge in middle and late gestation. The maternal administration of PolyI:C (5 mg/kg, i.v.) led to a marked increase in the serum protein levels of IL-1β, IL-6, IL-10, and TNF-α
    Figure Legend Snippet: Comparison of the maternal serum cytokine responses after immune challenge in middle and late gestation. The maternal administration of PolyI:C (5 mg/kg, i.v.) led to a marked increase in the serum protein levels of IL-1β, IL-6, IL-10, and TNF-α

    Techniques Used:

    36) Product Images from "A Large Polysaccharide Produced by Helicobacter hepaticus Induces an Anti-inflammatory Gene Signature in Macrophages"

    Article Title: A Large Polysaccharide Produced by Helicobacter hepaticus Induces an Anti-inflammatory Gene Signature in Macrophages

    Journal: Cell Host & Microbe

    doi: 10.1016/j.chom.2017.11.002

    H. hepaticus Produces a Large Soluble Polysaccharide-Inducing IL-10 Production in Macrophages (A–G) M-CSF-differentiated BMDMs were stimulated for 3 hr with different culture fractions of H. hepaticus . (A) mRNA and protein induction of the cytokines IL-10 and IL-6 after stimulation with control medium (TSB), H. hepaticus whole bacteria ( Hh ), or H. hepaticus -filtered cultured supernatant (SN Hh ). (B–D) Induction of (B) IL-10, (C) IL-6, and (D) IL-10/IL-6 ratio after 3 hr stimulation with SN Hh treated with enzymes and heat (SN Hh t), SN Hh t fractionated by size (SN Hh t > 30 kDa and SN Hh t
    Figure Legend Snippet: H. hepaticus Produces a Large Soluble Polysaccharide-Inducing IL-10 Production in Macrophages (A–G) M-CSF-differentiated BMDMs were stimulated for 3 hr with different culture fractions of H. hepaticus . (A) mRNA and protein induction of the cytokines IL-10 and IL-6 after stimulation with control medium (TSB), H. hepaticus whole bacteria ( Hh ), or H. hepaticus -filtered cultured supernatant (SN Hh ). (B–D) Induction of (B) IL-10, (C) IL-6, and (D) IL-10/IL-6 ratio after 3 hr stimulation with SN Hh treated with enzymes and heat (SN Hh t), SN Hh t fractionated by size (SN Hh t > 30 kDa and SN Hh t

    Techniques Used: Cell Culture

    MSK1/2 Is Essential to the Anti-inflammatory Properties of SN Hh t (A) Western blot showing the phosphorylation of CREB1 S133 and MSK1 T581 and the total CREB1 and MSK1 protein amounts in WT and Msk1/2 −/− BMDMs after 30 min stimulation with TSBt, SN Hh t, Pam3, or LPS UP. (B) mRNA levels of Il10 , Fosb , and Egr3 genes in BMDMs from WT or Msk1/2 −/− mice after 1 hr stimulation. (C) Induction of IL-10, IL-6, and TNFα; IL-10/IL-6; and IL-10/TNFα protein ratios in BMDMs from WT or Msk1/2 −/− mice after 10 hr stimulation. One of three independent experiments. Two-way ANOVA and Sidak’s and/or Tukey’s multiple comparisons tests, p
    Figure Legend Snippet: MSK1/2 Is Essential to the Anti-inflammatory Properties of SN Hh t (A) Western blot showing the phosphorylation of CREB1 S133 and MSK1 T581 and the total CREB1 and MSK1 protein amounts in WT and Msk1/2 −/− BMDMs after 30 min stimulation with TSBt, SN Hh t, Pam3, or LPS UP. (B) mRNA levels of Il10 , Fosb , and Egr3 genes in BMDMs from WT or Msk1/2 −/− mice after 1 hr stimulation. (C) Induction of IL-10, IL-6, and TNFα; IL-10/IL-6; and IL-10/TNFα protein ratios in BMDMs from WT or Msk1/2 −/− mice after 10 hr stimulation. One of three independent experiments. Two-way ANOVA and Sidak’s and/or Tukey’s multiple comparisons tests, p

    Techniques Used: Western Blot, Mouse Assay

    SN Hh t Is Sufficient to Induce IL-10 In Vivo (A) Frequency of IL-10 high cells among resident macrophages from caecum and colon LPL. SPF WT mice were infected with Hh or orally gavaged with TSBt or SN Hh t for 3 days. (B and C) Il10 , Il6 , and Tnf mRNA expression levels in the peritoneal cell fraction after 2 days (B) or Il10 and Il6 mRNA transcripts after 6 hr (C) challenge. Ligands (TSBt and SN Hh t, 200 μL; LPS UP and Pam3, 50 μg) were injected into the intraperitoneal cavity of SPF WT mice. Each symbol represents an individual mouse (two to three independent experiments). Pam3, Pam3CSK4. Mann-Whitney test, p
    Figure Legend Snippet: SN Hh t Is Sufficient to Induce IL-10 In Vivo (A) Frequency of IL-10 high cells among resident macrophages from caecum and colon LPL. SPF WT mice were infected with Hh or orally gavaged with TSBt or SN Hh t for 3 days. (B and C) Il10 , Il6 , and Tnf mRNA expression levels in the peritoneal cell fraction after 2 days (B) or Il10 and Il6 mRNA transcripts after 6 hr (C) challenge. Ligands (TSBt and SN Hh t, 200 μL; LPS UP and Pam3, 50 μg) were injected into the intraperitoneal cavity of SPF WT mice. Each symbol represents an individual mouse (two to three independent experiments). Pam3, Pam3CSK4. Mann-Whitney test, p

    Techniques Used: In Vivo, Mouse Assay, Infection, Expressing, Injection, MANN-WHITNEY

    H. hepaticus Induces IL-10 in Gut-Resident Macrophages (A–D) SPF WT mice were infected with Helicobacter hepaticus ( Hh ) for 3 or 5 days. (A) Experimental design. (B and C) FACS analysis of caecum and colon LPL after 3 days of infection with Hh . (B) Frequency of resident macrophages among total CD45 + cells. (C) Frequency of IL-10 high cells among resident macrophages, using an anti-mouse IL-10 antibody or its isotype control. (D) Expression level of Il10 , Il6 , and Tnf mRNA in caecum or colon tissue after 3 or 5 days of infection with Hh . Each symbol represents an individual mouse (two to three independent experiments). Mann-Whitney test (B and C) or one-way ANOVA and Tukey’s multiple comparisons test (D), p
    Figure Legend Snippet: H. hepaticus Induces IL-10 in Gut-Resident Macrophages (A–D) SPF WT mice were infected with Helicobacter hepaticus ( Hh ) for 3 or 5 days. (A) Experimental design. (B and C) FACS analysis of caecum and colon LPL after 3 days of infection with Hh . (B) Frequency of resident macrophages among total CD45 + cells. (C) Frequency of IL-10 high cells among resident macrophages, using an anti-mouse IL-10 antibody or its isotype control. (D) Expression level of Il10 , Il6 , and Tnf mRNA in caecum or colon tissue after 3 or 5 days of infection with Hh . Each symbol represents an individual mouse (two to three independent experiments). Mann-Whitney test (B and C) or one-way ANOVA and Tukey’s multiple comparisons test (D), p

    Techniques Used: Mouse Assay, Infection, FACS, Expressing, MANN-WHITNEY

    SN Hh t Stimulates CREB Phosphorylation and Induction of an Anti-inflammatory and Repair Gene Signature in Macrophages (A) Western blot showing the phosphorylation of CREB1 S133 and RelA S536 and the total CREB1 and RelA protein amounts in WT and Tlr2 −/− BMDMs after 30 min stimulation with TSBt, SN Hh t, Pam3, or LPS UP. β-actin used as a loading control. (B) mRNA levels of Il10 , Fosb , and Egr3 genes in BMDMs from conditional vav-cre CREB WT and CREB S133A KI mice after 1 hr stimulation with TSBt, Hh , SN Hh t, or Pam3. (C) Induction of IL-10, IL-6, and TNFα; IL-10/IL-6; and IL-10/TNFα protein ratios in BMDMs from conditional vav-cre CREB WT and CREB S133A KI mice after 10 hr stimulation with TSBt, Hh , SN Hh t, or Pam3. One of three independent experiments. Two-way ANOVA and Sidak’s and/or Tukey’s multiple comparisons tests, p
    Figure Legend Snippet: SN Hh t Stimulates CREB Phosphorylation and Induction of an Anti-inflammatory and Repair Gene Signature in Macrophages (A) Western blot showing the phosphorylation of CREB1 S133 and RelA S536 and the total CREB1 and RelA protein amounts in WT and Tlr2 −/− BMDMs after 30 min stimulation with TSBt, SN Hh t, Pam3, or LPS UP. β-actin used as a loading control. (B) mRNA levels of Il10 , Fosb , and Egr3 genes in BMDMs from conditional vav-cre CREB WT and CREB S133A KI mice after 1 hr stimulation with TSBt, Hh , SN Hh t, or Pam3. (C) Induction of IL-10, IL-6, and TNFα; IL-10/IL-6; and IL-10/TNFα protein ratios in BMDMs from conditional vav-cre CREB WT and CREB S133A KI mice after 10 hr stimulation with TSBt, Hh , SN Hh t, or Pam3. One of three independent experiments. Two-way ANOVA and Sidak’s and/or Tukey’s multiple comparisons tests, p

    Techniques Used: Western Blot, Mouse Assay

    37) Product Images from "Estrogen receptor-α in female skeletal muscle is not required for regulation of muscle insulin sensitivity and mitochondrial regulation"

    Article Title: Estrogen receptor-α in female skeletal muscle is not required for regulation of muscle insulin sensitivity and mitochondrial regulation

    Journal: Molecular Metabolism

    doi: 10.1016/j.molmet.2019.12.010

    Induced ablation of skeletal muscle ERα in adult female mice alters inflammatory markers in skeletal muscle, but not in systemic circulation . Key inflammatory cytokines and chemokines in  (A)  gastrocnemius (n = 7–9/group) and  (B)  blood serum of WT and ERαKO ism  mice after chronic HFD (n = 5–9/group). mRNA expression was measured by RT-qPCR and normalized to 18s rRNA. TNF-α = tumor necrosis factor-α; IL-6 = interleukin-6; IL-1β = interleukin-1β; CXCL2 = chemokine (C-X-C motif) ligand 2; CXCL1 = chemokine (C-X-C motif) ligand 1; CCL3 = chemokine (C–C motif) ligand 3; chemokine (C–C motif) ligand 2. Data are means ± SEM. *Main effect of diet,  p
    Figure Legend Snippet: Induced ablation of skeletal muscle ERα in adult female mice alters inflammatory markers in skeletal muscle, but not in systemic circulation . Key inflammatory cytokines and chemokines in (A) gastrocnemius (n = 7–9/group) and (B) blood serum of WT and ERαKO ism mice after chronic HFD (n = 5–9/group). mRNA expression was measured by RT-qPCR and normalized to 18s rRNA. TNF-α = tumor necrosis factor-α; IL-6 = interleukin-6; IL-1β = interleukin-1β; CXCL2 = chemokine (C-X-C motif) ligand 2; CXCL1 = chemokine (C-X-C motif) ligand 1; CCL3 = chemokine (C–C motif) ligand 3; chemokine (C–C motif) ligand 2. Data are means ± SEM. *Main effect of diet, p

    Techniques Used: Mouse Assay, Expressing, Quantitative RT-PCR

    38) Product Images from "Anti-Inflammatory Activity of Diterpenoids from Celastrus orbiculatus in Lipopolysaccharide-Stimulated RAW264.7 Cells"

    Article Title: Anti-Inflammatory Activity of Diterpenoids from Celastrus orbiculatus in Lipopolysaccharide-Stimulated RAW264.7 Cells

    Journal: Journal of Immunology Research

    doi: 10.1155/2020/7207354

    Compounds 1 and 3 downregulated proinflammatory mediators. (a–c) The mRNA expression levels of IL-1 β , IL-6, and TNF- α were measured using quantitative real-time PCR experiment, and these proinflammatory cytokines were significantly diminished by compounds 1 and 3 . Cells were preincubated for 2 h with compounds 1 and 3 at concentration of 5 and 10 μ M, respectively, and activated by LPS (1 μ g/mL) for 2 h. Results represent as mean ± SEM, and dexamethasone was used as a positive control. # p
    Figure Legend Snippet: Compounds 1 and 3 downregulated proinflammatory mediators. (a–c) The mRNA expression levels of IL-1 β , IL-6, and TNF- α were measured using quantitative real-time PCR experiment, and these proinflammatory cytokines were significantly diminished by compounds 1 and 3 . Cells were preincubated for 2 h with compounds 1 and 3 at concentration of 5 and 10 μ M, respectively, and activated by LPS (1 μ g/mL) for 2 h. Results represent as mean ± SEM, and dexamethasone was used as a positive control. # p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Positive Control

    39) Product Images from "Mesenchymal Stem Cell Therapy Modulates the Inflammatory Response in Experimental Traumatic Brain Injury"

    Article Title: Mesenchymal Stem Cell Therapy Modulates the Inflammatory Response in Experimental Traumatic Brain Injury

    Journal: Neurology Research International

    doi: 10.1155/2011/564089

    Relative expression of IL-6, TNF- α , IL-10, and IL-4 in acute and chronic models of motor cortex injury. (a) 24 hours and (b) 30 days after injury and MSC injection, RNA was extracted from motor cortex of control, injury without treatment and MSC treated mice. qPCR was performed to quantify the expression of inflammatory cytokines and relative expression was calculated in relation to HPRT. Data are expressed as mean of 2 −ΔΔCt ± SEM (control n = 3, injury no treatment n = 6, MSC treated n = 4). * P
    Figure Legend Snippet: Relative expression of IL-6, TNF- α , IL-10, and IL-4 in acute and chronic models of motor cortex injury. (a) 24 hours and (b) 30 days after injury and MSC injection, RNA was extracted from motor cortex of control, injury without treatment and MSC treated mice. qPCR was performed to quantify the expression of inflammatory cytokines and relative expression was calculated in relation to HPRT. Data are expressed as mean of 2 −ΔΔCt ± SEM (control n = 3, injury no treatment n = 6, MSC treated n = 4). * P

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

    40) Product Images from "IL-10-dependent Tr1 cells attenuate astrocyte activation and ameliorate chronic central nervous system inflammation"

    Article Title: IL-10-dependent Tr1 cells attenuate astrocyte activation and ameliorate chronic central nervous system inflammation

    Journal: Brain

    doi: 10.1093/brain/aww113

    Nasal anti-CD3 attenuates microglial activation. ( A–C ) Naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs during the chronic phase as in Fig. 1 A. ( A ) Heat map depicting mRNA expression, as detected by Nanostring nCounter analysis, in microglial cells isolated from naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs. Upper panels : Histogram presentation of normalized gene expression in each gene cluster. Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( B ) Quantitative PCR analysis of Il1b , Il6 , Nos2 , Cd40 , and Irf7 expression in microglial cells isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( C ) Mean normalized expression of genes associated with M1 or M2 phenotype in microglia ( Supplementary Table 6 ). Statistical analysis by Student’s t -test. ( D ) Primary microglia were pretreated for 1 h with IL-10 (100 ng/ml), or vehicle (PBS), followed by activation with GM-CSF (25 ng/ml) or left untreated. Quantitative PCR analysis of Il1b , Il6 , Il12b , Cd40 , H2aa , and TNF ; expression is presented as fold-change from untreated relative to Gapdh . Data from three independent experiments (mean and SEM). Statistical analysis by Student’s t- test. * P
    Figure Legend Snippet: Nasal anti-CD3 attenuates microglial activation. ( A–C ) Naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs during the chronic phase as in Fig. 1 A. ( A ) Heat map depicting mRNA expression, as detected by Nanostring nCounter analysis, in microglial cells isolated from naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs. Upper panels : Histogram presentation of normalized gene expression in each gene cluster. Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( B ) Quantitative PCR analysis of Il1b , Il6 , Nos2 , Cd40 , and Irf7 expression in microglial cells isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( C ) Mean normalized expression of genes associated with M1 or M2 phenotype in microglia ( Supplementary Table 6 ). Statistical analysis by Student’s t -test. ( D ) Primary microglia were pretreated for 1 h with IL-10 (100 ng/ml), or vehicle (PBS), followed by activation with GM-CSF (25 ng/ml) or left untreated. Quantitative PCR analysis of Il1b , Il6 , Il12b , Cd40 , H2aa , and TNF ; expression is presented as fold-change from untreated relative to Gapdh . Data from three independent experiments (mean and SEM). Statistical analysis by Student’s t- test. * P

    Techniques Used: Activation Assay, Mouse Assay, Expressing, Isolation, Real-time Polymerase Chain Reaction

    Nasal anti-CD3 suppresses astrocyte activation. ( A–E ) Naïve or EAE NOD mice treated with CD3-specific or isotype control mAbs during the chronic phase as in ( Fig. 1 A). ( A ) Heat map depicting mRNA expression, as detected by Nanostring nCounter analysis, in astrocytes isolated from naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs. Upper panels : Histogram presentation of normalized gene expression in each gene cluster. Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( B ) Quantitative PCR analysis of Ccl2 , Ccl5 , Il6 , Tlr2 , and Aqp4 expression in astrocytes isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( C ) Immunohistochemistry analysis of spinal cords for aquaporin 4 (AQP4) expression, and the presence of astrocytes (GFAP) on subsequent sections. Scale bar = 50 µm. Data are representative of two independent experiments with n = 6 mice/group. ( D ) Relative expression (to NOD naïve group) of genes associated with the control of demyelination in astrocytes isolated from EAE NOD mice treated with anti-CD3 or isotype control. Representative data of three independent experiments. Statistical analysis by Student’s t- test. ( E ) Quantitative PCR analysis of Mmp3 , Mmp9 and Vegfa expression in astrocytes isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( F ) Cultured astrocytes were pre-treated for 1 h with IL-10 (100 ng/ml), or vehicle (PBS), followed by activation with IL1β (20 ng/ml) or left untreated. Quantitative PCR analysis of Ccl2 , Ccl5 , Csf2 , Nos2 , Mmp3 , and Mmp9 ; expression is presented as fold change from untreated, relative to Gapdh . Data from five independent experiments (mean and SEM). Statistical analysis by two-way ANOVA, followed by Tukey post hoc analysis. * P
    Figure Legend Snippet: Nasal anti-CD3 suppresses astrocyte activation. ( A–E ) Naïve or EAE NOD mice treated with CD3-specific or isotype control mAbs during the chronic phase as in ( Fig. 1 A). ( A ) Heat map depicting mRNA expression, as detected by Nanostring nCounter analysis, in astrocytes isolated from naïve or EAE NOD mice treated with CD3 specific or isotype control mAbs. Upper panels : Histogram presentation of normalized gene expression in each gene cluster. Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( B ) Quantitative PCR analysis of Ccl2 , Ccl5 , Il6 , Tlr2 , and Aqp4 expression in astrocytes isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( C ) Immunohistochemistry analysis of spinal cords for aquaporin 4 (AQP4) expression, and the presence of astrocytes (GFAP) on subsequent sections. Scale bar = 50 µm. Data are representative of two independent experiments with n = 6 mice/group. ( D ) Relative expression (to NOD naïve group) of genes associated with the control of demyelination in astrocytes isolated from EAE NOD mice treated with anti-CD3 or isotype control. Representative data of three independent experiments. Statistical analysis by Student’s t- test. ( E ) Quantitative PCR analysis of Mmp3 , Mmp9 and Vegfa expression in astrocytes isolated from naïve and EAE NOD mice treated as in A ; expression is presented relative to Gapdh . Representative data of three independent experiments, statistical analysis by one-way ANOVA, followed by Tukey post hoc analysis. ( F ) Cultured astrocytes were pre-treated for 1 h with IL-10 (100 ng/ml), or vehicle (PBS), followed by activation with IL1β (20 ng/ml) or left untreated. Quantitative PCR analysis of Ccl2 , Ccl5 , Csf2 , Nos2 , Mmp3 , and Mmp9 ; expression is presented as fold change from untreated, relative to Gapdh . Data from five independent experiments (mean and SEM). Statistical analysis by two-way ANOVA, followed by Tukey post hoc analysis. * P

    Techniques Used: Activation Assay, Mouse Assay, Expressing, Isolation, Real-time Polymerase Chain Reaction, Immunohistochemistry, Cell Culture

    Related Articles

    Expressing:

    Article Title: Acute and chronic effects of exercise on mRNA expression in the skeletal muscle of two mouse models of peripheral artery disease
    Article Snippet: The RNA-containing supernatants were purified using an RNeasy Mini kit (Qiagen) and reverse-transcribed using a High-Capacity cDNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA) to obtain cDNA. .. The mRNA expression levels were then determined by TaqMan quantitative real-time reverse transcription PCR (qRT-PCR; Applied Biosystems, Foster City, CA), using the following predesigned primer/probe sets: Rplp0 , Mm99999223_gH; Ppargc1a (Pgc1a) , Mm01184322_m1; Il6 , Mm00446190_m1; Nr4a1 , Mm01300401_m1; Nr4a2 , Mm00443060_m1; Nr4a3 , Mm00450074_m1; Myf5 , Mm00435125_m1; Myogenin , Mm00446195_g1; Tmem8c (Myomaker) , Mm00481255_m1; Myh3 , Mm01332463_m1; Gpr56 , Mm00817704_m1; and Col3a1 , Mm01254476_m1. .. Each expression value was normalized to the expression levels of Rplp0 and calculated relative to the pre-exercise group for the acute phase study and to the sham-operated sedentary group for the chronic phase study.

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages
    Article Snippet: This modality of indirect co-culture allowed us to recover isolated adipocytes. cDNA synthesis was carried out in a volume of 20 μL using 2 μg total RNA, M-MLV reverse transcriptase and 0.5 mM random primers (Invitrogene Life Technologies). .. Subsequently, cDNA was amplified by quantitative real time polymerase chain reaction (qPCR), using Taqman probe-based gene expression assays (Applied Biosystems) as follows: (MCP-1) Mm00441242_m1; (IL-6) Mm00446190_m1; (TNF-α) Mm00443260_g1; (actin-ß) Mm00607939_s1. .. Amplification was performed in a total reaction volume of 10 μL containing 100 ng cDNA, 3.5 μL H2 O, 0.5 μL of the Taqman probe-based gene expression assay and 1X Taqman gene expression Master Mix (Applied Biosystems) in a Rotor Gene fluorescence thermal cycler (Qiagen, Limburg, NL) for 45 cycles.

    Article Title: IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content
    Article Snippet: SuperScript II Reverse Transcriptase, oligo(dT) and dNTP were products from Invitrogen (Burlington, ON). .. Taqman Gene Expression Assays for mouse β actin (4352933E), PGC-1α (CAT# Mm01208835_m1), PGC-1β (CAT# Mm00504720_m1), PRC (CAT# Mm00521078_m1), COXIV (CAT# Mm01250094_m1), CPT-1 (CAT # Mm01308166_m1) and IL-6 (CAT# Mm00446190_m1) were from Applied Biosytems (Foster City, CA). ..

    Article Title: Cardiac Function Remains Impaired Despite Reversible Cardiac Remodeling after Acute Experimental Viral Myocarditis
    Article Snippet: Relative quantification of RNA expression was performed with a 7900 TaqMan system (Applied Biosystems, Germany). .. To determine RNA expression levels of various genes, real-time PCR was carried out using 5 μ L of gene expression master mix (Life technologies, Germany) and 0.5 μ L of the gene expression assay for Mcp-1 (Ccl2) (Mm99999056_m1), Rantes (Ccl5) (Mm01302428_m1), Mcp-3 (Ccl7) (Mm00443113_m1), Ip-10 (Cxcl10) (Mm99999072_m1), Cxcl13 (Mm00444534_m1), Il-6 (Mm00446190_m1), Il-23a (Mm00518984_m1), Tnf-α (Mm00443258_m1), Cd3 (Mm01179194_m1), Cd4 (Mm00442754_m1), Cd8 (Mm01182107_g1), Cd19 (Mm00515420_m1), Col1a1 (Mm01302043_g1), Col3a1 (Mm00802331_m1), Ctgf (Mm00515790_g1), Tgf-β1 (Mm00441724_m1), Timp-1 (Mm00441818_m1), or Mmp13 (Mm01168712_m1) purchased from Life Technologies. ..

    Article Title: Obesity enhances sepsis induced liver inflammation and injury in mice
    Article Snippet: The mRNA was reversely transcribed to cDNA using the high-capacity cDNA reverse transcription kit (Applied Biosystems, Grand Island, NY). .. Comparative qPCR was performed using TaqMan Gene expression master mix (Applied Biosystems) with the following primers and probes: GAPDH (Mm99999915_gl), IL-6 (Mm00446190_ml), TNFα (Mm00443258_ml), AdipoR2 (Mm11184032_m1) and PPARγ (Mm01184332-m1). .. The reaction was performed and analyzed using a quantitative PCR system (QuantStudio-6Flex machine, Applied Biosystems).

    Article Title: Attenuated viral hepatitis in Trem1−/− mice is associated with reduced inflammatory activity of neutrophils
    Article Snippet: Gene expression was normalized to the peptidylprolyl isomerase A (PPIA ) housekeeper mRNA. .. TaqMan assay-on-Demand primer/probe sets (Life Technologies) were used for the detection of gene expression for TREM1: Mm01278455_m1, DAP12: Mm00449152_m1, PPIA: Mm02342429_g1, CCL2: Mm00441242_m1, TNF-α: Mm00443258_m1, IL-1β: Mm00434228_m1, IL-6: Mm00446190_m1, MPO: Mm01298424_m1, CXCL1: Mm04207460_m1, CXCL2: Mm00436450_m1, CXCL5: Mm00436451_g1, IFN-α: Mm03030145_gH and IFN-γ: Mm01168134_m1. .. LCMV Z RNA expression was quantified by using the LightCycler FastStart DNA Master SYBR Green I hot start reaction mix (Roche) on a LightCycler© 1.5 system (Roche) with custom-made primer pairs (LCMV_Z_fwd: 5′-CAGACACCACCTATCTTGG-3′ and LCMV_Z_rev: 3′ ACCTTCAG TTTGGTTGGC-5′).

    Polymerase Chain Reaction:

    Article Title: Acute and chronic effects of exercise on mRNA expression in the skeletal muscle of two mouse models of peripheral artery disease
    Article Snippet: The RNA-containing supernatants were purified using an RNeasy Mini kit (Qiagen) and reverse-transcribed using a High-Capacity cDNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA) to obtain cDNA. .. The mRNA expression levels were then determined by TaqMan quantitative real-time reverse transcription PCR (qRT-PCR; Applied Biosystems, Foster City, CA), using the following predesigned primer/probe sets: Rplp0 , Mm99999223_gH; Ppargc1a (Pgc1a) , Mm01184322_m1; Il6 , Mm00446190_m1; Nr4a1 , Mm01300401_m1; Nr4a2 , Mm00443060_m1; Nr4a3 , Mm00450074_m1; Myf5 , Mm00435125_m1; Myogenin , Mm00446195_g1; Tmem8c (Myomaker) , Mm00481255_m1; Myh3 , Mm01332463_m1; Gpr56 , Mm00817704_m1; and Col3a1 , Mm01254476_m1. .. Each expression value was normalized to the expression levels of Rplp0 and calculated relative to the pre-exercise group for the acute phase study and to the sham-operated sedentary group for the chronic phase study.

    Article Title: Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury
    Article Snippet: After digestion with DNase I, 2 μg RNA was reverse transcribed with the Applied Biosystems reverse transcriptase kit (Applied Biosystems, CA, USA). .. The quality of cDNA was assessed by the ratio of the absorbance at 260 nm and 280 nm using an Agilent 2100 Bioanalyzer (Agilent Bioanalyzer, CA, USA). cDNA was then amplified by PCR in an iCycler Thermal Cycler (Bio-Rad, CA, USA) with SYBR Green (Applied Biosystems, CA, USA) and specific primers for Kim1 (Mm01291075_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Timp1 (Mm01341361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Ngal (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Il6 (Mm00446190_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Tnfa (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Nfkb1 (Mm00476361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Gapdh (Mm99999915_g1, Thermo Fisher Scientific, Santa Clara, CA, USA), and 18S (Mm02601777_g1, Thermo Fisher Scientific, Santa Clara, CA, USA); the latter two were used as housekeeping genes. .. The expression of marker genes was normalized to the endogenous Gapdh expression level and calculated with the 2−ΔΔCt formula in % Gapdh expression [ ].

    Quantitative RT-PCR:

    Article Title: Acute and chronic effects of exercise on mRNA expression in the skeletal muscle of two mouse models of peripheral artery disease
    Article Snippet: The RNA-containing supernatants were purified using an RNeasy Mini kit (Qiagen) and reverse-transcribed using a High-Capacity cDNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA) to obtain cDNA. .. The mRNA expression levels were then determined by TaqMan quantitative real-time reverse transcription PCR (qRT-PCR; Applied Biosystems, Foster City, CA), using the following predesigned primer/probe sets: Rplp0 , Mm99999223_gH; Ppargc1a (Pgc1a) , Mm01184322_m1; Il6 , Mm00446190_m1; Nr4a1 , Mm01300401_m1; Nr4a2 , Mm00443060_m1; Nr4a3 , Mm00450074_m1; Myf5 , Mm00435125_m1; Myogenin , Mm00446195_g1; Tmem8c (Myomaker) , Mm00481255_m1; Myh3 , Mm01332463_m1; Gpr56 , Mm00817704_m1; and Col3a1 , Mm01254476_m1. .. Each expression value was normalized to the expression levels of Rplp0 and calculated relative to the pre-exercise group for the acute phase study and to the sham-operated sedentary group for the chronic phase study.

    Amplification:

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages
    Article Snippet: This modality of indirect co-culture allowed us to recover isolated adipocytes. cDNA synthesis was carried out in a volume of 20 μL using 2 μg total RNA, M-MLV reverse transcriptase and 0.5 mM random primers (Invitrogene Life Technologies). .. Subsequently, cDNA was amplified by quantitative real time polymerase chain reaction (qPCR), using Taqman probe-based gene expression assays (Applied Biosystems) as follows: (MCP-1) Mm00441242_m1; (IL-6) Mm00446190_m1; (TNF-α) Mm00443260_g1; (actin-ß) Mm00607939_s1. .. Amplification was performed in a total reaction volume of 10 μL containing 100 ng cDNA, 3.5 μL H2 O, 0.5 μL of the Taqman probe-based gene expression assay and 1X Taqman gene expression Master Mix (Applied Biosystems) in a Rotor Gene fluorescence thermal cycler (Qiagen, Limburg, NL) for 45 cycles.

    Article Title: Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury
    Article Snippet: After digestion with DNase I, 2 μg RNA was reverse transcribed with the Applied Biosystems reverse transcriptase kit (Applied Biosystems, CA, USA). .. The quality of cDNA was assessed by the ratio of the absorbance at 260 nm and 280 nm using an Agilent 2100 Bioanalyzer (Agilent Bioanalyzer, CA, USA). cDNA was then amplified by PCR in an iCycler Thermal Cycler (Bio-Rad, CA, USA) with SYBR Green (Applied Biosystems, CA, USA) and specific primers for Kim1 (Mm01291075_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Timp1 (Mm01341361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Ngal (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Il6 (Mm00446190_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Tnfa (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Nfkb1 (Mm00476361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Gapdh (Mm99999915_g1, Thermo Fisher Scientific, Santa Clara, CA, USA), and 18S (Mm02601777_g1, Thermo Fisher Scientific, Santa Clara, CA, USA); the latter two were used as housekeeping genes. .. The expression of marker genes was normalized to the endogenous Gapdh expression level and calculated with the 2−ΔΔCt formula in % Gapdh expression [ ].

    Real-time Polymerase Chain Reaction:

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages
    Article Snippet: This modality of indirect co-culture allowed us to recover isolated adipocytes. cDNA synthesis was carried out in a volume of 20 μL using 2 μg total RNA, M-MLV reverse transcriptase and 0.5 mM random primers (Invitrogene Life Technologies). .. Subsequently, cDNA was amplified by quantitative real time polymerase chain reaction (qPCR), using Taqman probe-based gene expression assays (Applied Biosystems) as follows: (MCP-1) Mm00441242_m1; (IL-6) Mm00446190_m1; (TNF-α) Mm00443260_g1; (actin-ß) Mm00607939_s1. .. Amplification was performed in a total reaction volume of 10 μL containing 100 ng cDNA, 3.5 μL H2 O, 0.5 μL of the Taqman probe-based gene expression assay and 1X Taqman gene expression Master Mix (Applied Biosystems) in a Rotor Gene fluorescence thermal cycler (Qiagen, Limburg, NL) for 45 cycles.

    Article Title: Cardiac Function Remains Impaired Despite Reversible Cardiac Remodeling after Acute Experimental Viral Myocarditis
    Article Snippet: Relative quantification of RNA expression was performed with a 7900 TaqMan system (Applied Biosystems, Germany). .. To determine RNA expression levels of various genes, real-time PCR was carried out using 5 μ L of gene expression master mix (Life technologies, Germany) and 0.5 μ L of the gene expression assay for Mcp-1 (Ccl2) (Mm99999056_m1), Rantes (Ccl5) (Mm01302428_m1), Mcp-3 (Ccl7) (Mm00443113_m1), Ip-10 (Cxcl10) (Mm99999072_m1), Cxcl13 (Mm00444534_m1), Il-6 (Mm00446190_m1), Il-23a (Mm00518984_m1), Tnf-α (Mm00443258_m1), Cd3 (Mm01179194_m1), Cd4 (Mm00442754_m1), Cd8 (Mm01182107_g1), Cd19 (Mm00515420_m1), Col1a1 (Mm01302043_g1), Col3a1 (Mm00802331_m1), Ctgf (Mm00515790_g1), Tgf-β1 (Mm00441724_m1), Timp-1 (Mm00441818_m1), or Mmp13 (Mm01168712_m1) purchased from Life Technologies. ..

    Article Title: Obesity enhances sepsis induced liver inflammation and injury in mice
    Article Snippet: The mRNA was reversely transcribed to cDNA using the high-capacity cDNA reverse transcription kit (Applied Biosystems, Grand Island, NY). .. Comparative qPCR was performed using TaqMan Gene expression master mix (Applied Biosystems) with the following primers and probes: GAPDH (Mm99999915_gl), IL-6 (Mm00446190_ml), TNFα (Mm00443258_ml), AdipoR2 (Mm11184032_m1) and PPARγ (Mm01184332-m1). .. The reaction was performed and analyzed using a quantitative PCR system (QuantStudio-6Flex machine, Applied Biosystems).

    Pyrolysis Gas Chromatography:

    Article Title: IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content
    Article Snippet: SuperScript II Reverse Transcriptase, oligo(dT) and dNTP were products from Invitrogen (Burlington, ON). .. Taqman Gene Expression Assays for mouse β actin (4352933E), PGC-1α (CAT# Mm01208835_m1), PGC-1β (CAT# Mm00504720_m1), PRC (CAT# Mm00521078_m1), COXIV (CAT# Mm01250094_m1), CPT-1 (CAT # Mm01308166_m1) and IL-6 (CAT# Mm00446190_m1) were from Applied Biosytems (Foster City, CA). ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content
    Article Snippet: SuperScript II Reverse Transcriptase, oligo(dT) and dNTP were products from Invitrogen (Burlington, ON). .. Taqman Gene Expression Assays for mouse β actin (4352933E), PGC-1α (CAT# Mm01208835_m1), PGC-1β (CAT# Mm00504720_m1), PRC (CAT# Mm00521078_m1), COXIV (CAT# Mm01250094_m1), CPT-1 (CAT # Mm01308166_m1) and IL-6 (CAT# Mm00446190_m1) were from Applied Biosytems (Foster City, CA). ..

    Cycling Probe Technology:

    Article Title: IL-6 Is Not Necessary for the Regulation of Adipose Tissue Mitochondrial Content
    Article Snippet: SuperScript II Reverse Transcriptase, oligo(dT) and dNTP were products from Invitrogen (Burlington, ON). .. Taqman Gene Expression Assays for mouse β actin (4352933E), PGC-1α (CAT# Mm01208835_m1), PGC-1β (CAT# Mm00504720_m1), PRC (CAT# Mm00521078_m1), COXIV (CAT# Mm01250094_m1), CPT-1 (CAT # Mm01308166_m1) and IL-6 (CAT# Mm00446190_m1) were from Applied Biosytems (Foster City, CA). ..

    RNA Expression:

    Article Title: Cardiac Function Remains Impaired Despite Reversible Cardiac Remodeling after Acute Experimental Viral Myocarditis
    Article Snippet: Relative quantification of RNA expression was performed with a 7900 TaqMan system (Applied Biosystems, Germany). .. To determine RNA expression levels of various genes, real-time PCR was carried out using 5 μ L of gene expression master mix (Life technologies, Germany) and 0.5 μ L of the gene expression assay for Mcp-1 (Ccl2) (Mm99999056_m1), Rantes (Ccl5) (Mm01302428_m1), Mcp-3 (Ccl7) (Mm00443113_m1), Ip-10 (Cxcl10) (Mm99999072_m1), Cxcl13 (Mm00444534_m1), Il-6 (Mm00446190_m1), Il-23a (Mm00518984_m1), Tnf-α (Mm00443258_m1), Cd3 (Mm01179194_m1), Cd4 (Mm00442754_m1), Cd8 (Mm01182107_g1), Cd19 (Mm00515420_m1), Col1a1 (Mm01302043_g1), Col3a1 (Mm00802331_m1), Ctgf (Mm00515790_g1), Tgf-β1 (Mm00441724_m1), Timp-1 (Mm00441818_m1), or Mmp13 (Mm01168712_m1) purchased from Life Technologies. ..

    SYBR Green Assay:

    Article Title: Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury
    Article Snippet: After digestion with DNase I, 2 μg RNA was reverse transcribed with the Applied Biosystems reverse transcriptase kit (Applied Biosystems, CA, USA). .. The quality of cDNA was assessed by the ratio of the absorbance at 260 nm and 280 nm using an Agilent 2100 Bioanalyzer (Agilent Bioanalyzer, CA, USA). cDNA was then amplified by PCR in an iCycler Thermal Cycler (Bio-Rad, CA, USA) with SYBR Green (Applied Biosystems, CA, USA) and specific primers for Kim1 (Mm01291075_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Timp1 (Mm01341361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Ngal (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Il6 (Mm00446190_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Tnfa (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Nfkb1 (Mm00476361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Gapdh (Mm99999915_g1, Thermo Fisher Scientific, Santa Clara, CA, USA), and 18S (Mm02601777_g1, Thermo Fisher Scientific, Santa Clara, CA, USA); the latter two were used as housekeeping genes. .. The expression of marker genes was normalized to the endogenous Gapdh expression level and calculated with the 2−ΔΔCt formula in % Gapdh expression [ ].

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher gene exp retnla mm00445109 m1
    The cellular senescent marker, <t>cyclin-dependent</t> kinase <t>inhibitor</t> 2A ( Cdkn2a ), increased in the LV with age. Cdkn2a expression was similarly increased with age in the LVs of both WT (solid bars) and SPARC-null (open bars) mice. Expression was normalized
    Gene Exp Retnla Mm00445109 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp retnla mm00445109 m1/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gene exp retnla mm00445109 m1 - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp il6 mm00446190 m1
    Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) <t>IL-6</t> levels after CLP. *p
    Gene Exp Il6 Mm00446190 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 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 1 article reviews
    Price from $9.99 to $1999.99
    gene exp il6 mm00446190 m1 - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    The cellular senescent marker, cyclin-dependent kinase inhibitor 2A ( Cdkn2a ), increased in the LV with age. Cdkn2a expression was similarly increased with age in the LVs of both WT (solid bars) and SPARC-null (open bars) mice. Expression was normalized

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Secreted protein acidic and rich in cysteine facilitates age-related cardiac inflammation and macrophage M1 polarization

    doi: 10.1152/ajpcell.00402.2014

    Figure Lengend Snippet: The cellular senescent marker, cyclin-dependent kinase inhibitor 2A ( Cdkn2a ), increased in the LV with age. Cdkn2a expression was similarly increased with age in the LVs of both WT (solid bars) and SPARC-null (open bars) mice. Expression was normalized

    Article Snippet: For cyclin-dependent kinase inhibitor 2A ( Cdkn2a ), chemokine (C-X3-C motif) ligand 1 ( Cx3cl1 ), chemokine (C-X-C motif) ligand 5 ( Cxcl5 ), interleukin-6 ( IL-6 ), tumor necrosis factor-α ( Tnfα ), and Fizz-1 expression, Taqman gene expression assays were performed using specific primers (Mm00494449_m1, Mm00436454_m1, Mm00436451_g1, Mm00446190_m1, Mm00443260_g1, and Mm00445109_m1; Applied Biosytems).

    Techniques: Marker, Expressing, Mouse Assay

    Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) IL-6 levels after CLP. *p

    Journal: Obesity (Silver Spring, Md.)

    Article Title: Obesity enhances sepsis induced liver inflammation and injury in mice

    doi: 10.1002/oby.21504

    Figure Lengend Snippet: Obesity alters cytokines after polymicrobial sepsis. Mice were randomized to a HFD or ND for 6–7wks. Polymicrobial sepsis was induced by CLP after diet intervention. Plasma (A) IL-17a (B) IL-23 (C) TNFα and (D) IL-6 levels after CLP. *p

    Article Snippet: Comparative qPCR was performed using TaqMan Gene expression master mix (Applied Biosystems) with the following primers and probes: GAPDH (Mm99999915_gl), IL-6 (Mm00446190_ml), TNFα (Mm00443258_ml), AdipoR2 (Mm11184032_m1) and PPARγ (Mm01184332-m1).

    Techniques: Mouse Assay

    The expression of (A) Adiponectin Receptor 2 (B) IL-6 (C) TNFα and (D) PPARγ in WAT were measured by RT-qPCR after CLP. *p

    Journal: Obesity (Silver Spring, Md.)

    Article Title: Obesity enhances sepsis induced liver inflammation and injury in mice

    doi: 10.1002/oby.21504

    Figure Lengend Snippet: The expression of (A) Adiponectin Receptor 2 (B) IL-6 (C) TNFα and (D) PPARγ in WAT were measured by RT-qPCR after CLP. *p

    Article Snippet: Comparative qPCR was performed using TaqMan Gene expression master mix (Applied Biosystems) with the following primers and probes: GAPDH (Mm99999915_gl), IL-6 (Mm00446190_ml), TNFα (Mm00443258_ml), AdipoR2 (Mm11184032_m1) and PPARγ (Mm01184332-m1).

    Techniques: Expressing, Quantitative RT-PCR

    3-3- diindolylmethane (DIM) effect on MCP-1, IL-6 and TNF-α production in supernatants of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW 264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane, treated with 20 µM, 40 µM, and 60 µM DIM during 24 h followed by 100 nM insulin for 20 min. Determinations were carried out in triplicate using individual enzyme-linked immunosorbent assay and expressed as the mean value ± SD *p

    Journal: Current Medicinal Chemistry

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages

    doi: 10.2174/1573406413666170922095011

    Figure Lengend Snippet: 3-3- diindolylmethane (DIM) effect on MCP-1, IL-6 and TNF-α production in supernatants of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW 264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane, treated with 20 µM, 40 µM, and 60 µM DIM during 24 h followed by 100 nM insulin for 20 min. Determinations were carried out in triplicate using individual enzyme-linked immunosorbent assay and expressed as the mean value ± SD *p

    Article Snippet: Subsequently, cDNA was amplified by quantitative real time polymerase chain reaction (qPCR), using Taqman probe-based gene expression assays (Applied Biosystems) as follows: (MCP-1) Mm00441242_m1; (IL-6) Mm00446190_m1; (TNF-α) Mm00443260_g1; (actin-ß) Mm00607939_s1.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    3-3-diindolylmethane (DIM) effect on MCP-1, IL-6, and TNF -α mRNA relative expression of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane treated with 20 µM, 40 µM, and 60 µM DIM during 24 h, followed by 100 nM insulin for 20 min. Determinations were performed by qPCR. Results were normalized based on the expression of actin-ß gene and analyzed by the comparative relative expression CT (2 -∆∆C T ) method [ 31 ] *p

    Journal: Current Medicinal Chemistry

    Article Title: IRS-1 pY612 and Akt-1/PKB pT308 Phosphorylation and Anti-inflammatory Effect of Diindolylmethane in Adipocytes Cocultured with Macrophages

    doi: 10.2174/1573406413666170922095011

    Figure Lengend Snippet: 3-3-diindolylmethane (DIM) effect on MCP-1, IL-6, and TNF -α mRNA relative expression of 3T3-L1 adipocytes (1 x10 5 ) co-cultured with RAW264.7 macrophages (5 x10 4 ) in a transwell system with a 0.4 µm porous membrane treated with 20 µM, 40 µM, and 60 µM DIM during 24 h, followed by 100 nM insulin for 20 min. Determinations were performed by qPCR. Results were normalized based on the expression of actin-ß gene and analyzed by the comparative relative expression CT (2 -∆∆C T ) method [ 31 ] *p

    Article Snippet: Subsequently, cDNA was amplified by quantitative real time polymerase chain reaction (qPCR), using Taqman probe-based gene expression assays (Applied Biosystems) as follows: (MCP-1) Mm00441242_m1; (IL-6) Mm00446190_m1; (TNF-α) Mm00443260_g1; (actin-ß) Mm00607939_s1.

    Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    Inflammatory cytokines. a Immunohistochemistry staining for inflammatory markers TNF-α and IL-6 in kidney tissue. b Western blot on kidney tissue measuring inflammatory markers TNF-α and NF-κB (left), alongside their quantification (right). c Quantitative real-time PCR on kidney tissue measuring inflammatory markers Nfkb , Il6 , and Tnfa . d ELISA measurement of blood serum concentrations of cytokines IL-1β, IL-6, and TNF-α. Each group has n = 5 mice. Significant difference a p

    Journal: Stem Cell Research & Therapy

    Article Title: Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury

    doi: 10.1186/s13287-020-01922-1

    Figure Lengend Snippet: Inflammatory cytokines. a Immunohistochemistry staining for inflammatory markers TNF-α and IL-6 in kidney tissue. b Western blot on kidney tissue measuring inflammatory markers TNF-α and NF-κB (left), alongside their quantification (right). c Quantitative real-time PCR on kidney tissue measuring inflammatory markers Nfkb , Il6 , and Tnfa . d ELISA measurement of blood serum concentrations of cytokines IL-1β, IL-6, and TNF-α. Each group has n = 5 mice. Significant difference a p

    Article Snippet: The quality of cDNA was assessed by the ratio of the absorbance at 260 nm and 280 nm using an Agilent 2100 Bioanalyzer (Agilent Bioanalyzer, CA, USA). cDNA was then amplified by PCR in an iCycler Thermal Cycler (Bio-Rad, CA, USA) with SYBR Green (Applied Biosystems, CA, USA) and specific primers for Kim1 (Mm01291075_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Timp1 (Mm01341361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Ngal (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Il6 (Mm00446190_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Tnfa (Mm00443258_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Nfkb1 (Mm00476361_m1, Thermo Fisher Scientific, Santa Clara, CA, USA), Gapdh (Mm99999915_g1, Thermo Fisher Scientific, Santa Clara, CA, USA), and 18S (Mm02601777_g1, Thermo Fisher Scientific, Santa Clara, CA, USA); the latter two were used as housekeeping genes.

    Techniques: Immunohistochemistry, Staining, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Mouse Assay