lps-induced Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore lps induced endotoxin shock
    Lps Induced Endotoxin Shock, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced endotoxin shock/product/Millipore
    Average 99 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    lps induced endotoxin shock - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    Takeda k commensal microbiota induce lps hyporesponsiveness
    K Commensal Microbiota Induce Lps Hyporesponsiveness, supplied by Takeda, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/k commensal microbiota induce lps hyporesponsiveness/product/Takeda
    Average 91 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    k commensal microbiota induce lps hyporesponsiveness - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    85
    Novoprotein lps induced mir 147 expression
    <t>miR-147</t> is a negative regulator of macrophage inflammatory responses. ( A–C ) miR-147 mimics attenuate <t>LPS,</t> PAM3CSK4, and poly(I:C) induced inflammatory response in primary macrophages. Peritoneal macrophages were transfected with 40 nM control
    Lps Induced Mir 147 Expression, supplied by Novoprotein, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced mir 147 expression/product/Novoprotein
    Average 85 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    lps induced mir 147 expression - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    86
    DuPont de Nemours e excessive in vitro bacterial lipopolysaccharide induced production
    <t>miR-147</t> is a negative regulator of macrophage inflammatory responses. ( A–C ) miR-147 mimics attenuate <t>LPS,</t> PAM3CSK4, and poly(I:C) induced inflammatory response in primary macrophages. Peritoneal macrophages were transfected with 40 nM control
    E Excessive In Vitro Bacterial Lipopolysaccharide Induced Production, supplied by DuPont de Nemours, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e excessive in vitro bacterial lipopolysaccharide induced production/product/DuPont de Nemours
    Average 86 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    e excessive in vitro bacterial lipopolysaccharide induced production - by Bioz Stars, 2020-09
    86/100 stars
      Buy from Supplier

    85
    SLIT2 LTD lps induced endothelial inflammation
    The anti-inflammatory effect of <t>Slit2</t> was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lps Induced Endothelial Inflammation, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced endothelial inflammation/product/SLIT2 LTD
    Average 85 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    lps induced endothelial inflammation - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    99
    Millipore lps induced tnfalpha synthesis
    The anti-inflammatory effect of <t>Slit2</t> was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lps Induced Tnfalpha Synthesis, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced tnfalpha synthesis/product/Millipore
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    lps induced tnfalpha synthesis - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    85
    SLIT2 LTD lipopolysaccharide lps induced secretion
    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting <t>HUVECs</t> were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lipopolysaccharide Lps Induced Secretion, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipopolysaccharide lps induced secretion/product/SLIT2 LTD
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    lipopolysaccharide lps induced secretion - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    85
    SLIT2 LTD lps induced anti inflammatory cytokines
    Slit2 inhibits <t>LPS-induced</t> Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. <t>Cytokines</t> secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p
    Lps Induced Anti Inflammatory Cytokines, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced anti inflammatory cytokines/product/SLIT2 LTD
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    lps induced anti inflammatory cytokines - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    88
    SLIT2 LTD lps induced inflammation
    The anti-inflammatory effect of <t>Slit2</t> was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lps Induced Inflammation, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced inflammation/product/SLIT2 LTD
    Average 88 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    lps induced inflammation - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    93
    SLIT2 LTD lps induced lung inflammation
    Effects of lipopolysaccharide <t>(LPS)</t> on <t>Slit2</t> expression. ( A ) Lung inflammation induced by the LPS treatment. ( B ) Expression of the Slit2-exon15 isoforms in the lungs and brains of mice treated with an intranasal administration of LPS. ( C ) Total expression level of Slit2 in the lungs relative to 18S rRNA. ( D ) Total expression level of Slit2 in the brain relative to 18S rRNA. (ΔCT = CT slit2 − CT 18SrRNA ; CT: cycle threshold). Higher ΔCT means a lower expression of Slit2. * p
    Lps Induced Lung Inflammation, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced lung inflammation/product/SLIT2 LTD
    Average 93 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    lps induced lung inflammation - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    85
    SLIT2 LTD lps induced significant downregulated slit2
    The anti-inflammatory effect of <t>Slit2</t> was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lps Induced Significant Downregulated Slit2, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced significant downregulated slit2/product/SLIT2 LTD
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    lps induced significant downregulated slit2 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    99
    Millipore lps induced
    The anti-inflammatory effect of <t>Slit2</t> was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lps Induced, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced/product/Millipore
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    lps induced - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    85
    SLIT2 LTD treatment inhibited lps induced pyk2 activation
    Slit2 inhibits <t>LPS-induced</t> <t>Pyk2</t> activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p
    Treatment Inhibited Lps Induced Pyk2 Activation, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/treatment inhibited lps induced pyk2 activation/product/SLIT2 LTD
    Average 85 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    treatment inhibited lps induced pyk2 activation - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    85
    SLIT2 LTD enhanced lps induced gm csf expression
    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, <t>GM-CSF</t> mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Enhanced Lps Induced Gm Csf Expression, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enhanced lps induced gm csf expression/product/SLIT2 LTD
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    enhanced lps induced gm csf expression - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    99
    Millipore 2 2 2 mouse intratracheal lps induced ali model e coli o55 b5 lps
    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, <t>GM-CSF</t> mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    2 2 2 Mouse Intratracheal Lps Induced Ali Model E Coli O55 B5 Lps, supplied by Millipore, 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/2 2 2 mouse intratracheal lps induced ali model e coli o55 b5 lps/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    2 2 2 mouse intratracheal lps induced ali model e coli o55 b5 lps - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    88
    Sirtris lipopolysaccharide lps induced cellular
    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, <t>GM-CSF</t> mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after <t>LPS</t> (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p
    Lipopolysaccharide Lps Induced Cellular, supplied by Sirtris, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipopolysaccharide lps induced cellular/product/Sirtris
    Average 88 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    lipopolysaccharide lps induced cellular - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    99
    Millipore lipopolysaccharide lps calbiochem induced tnf α release
    The anti-inflammatory effects of apoptotic cells are mediated through activation of JAK/STAT3 signalling Apoptotic neutrophil culture supernatants (apopt. SN) suppress <t>LPS-induced</t> <t>TNF-α</t> release. Monocytes were pre-treated for 3 h with apopt. SN and subsequently stimulated with LPS for 6 h. TNF-α contents in the culture media were measured by TNF-α ELISA. Activation of STAT3. Monocytes were treated with culture supernatants from either fresh or apoptotic cells for 1 h. STAT3 activation was detected by immunoblotting of cell lysates with the anti-pTYR705-STAT3 antibody. Total STAT3 levels were visualized by reprobing the membrane using a STAT3 antibody. Involvement of JAK in the dampening of acute <t>TNF-α</t> release by apoptotic cell supernatants. Monocytes were treated with apopt. SN for 3 h with or without a prior incubation with JAK inhibitor 1 for 15 min. Monocytes were then stimulated with LPS for 6 h and TNF-α levels in the culture media were measured by ELISA. STAT3 dependence of the inhibitory effect of apoptotic cell supernatants. Monocytes were pre-treated with the specific STAT3 Inhibitor VI for 30 min prior to treatment with the apoptotic neutrophil supernatant for 3 h and the LPS stimulation for 6 h. ELISA data are presented as the mean ± SEM of four independent experiments. The significance of the differences was evaluated by one-way ANOVA followed by Bonferroni's Multiple Comparison Test. p -values
    Lipopolysaccharide Lps Calbiochem Induced Tnf α Release, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipopolysaccharide lps calbiochem induced tnf α release/product/Millipore
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    lipopolysaccharide lps calbiochem induced tnf α release - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    SLIT2 LTD lps induced acute inflammation
    The anti-inflammatory effects of apoptotic cells are mediated through activation of JAK/STAT3 signalling Apoptotic neutrophil culture supernatants (apopt. SN) suppress <t>LPS-induced</t> <t>TNF-α</t> release. Monocytes were pre-treated for 3 h with apopt. SN and subsequently stimulated with LPS for 6 h. TNF-α contents in the culture media were measured by TNF-α ELISA. Activation of STAT3. Monocytes were treated with culture supernatants from either fresh or apoptotic cells for 1 h. STAT3 activation was detected by immunoblotting of cell lysates with the anti-pTYR705-STAT3 antibody. Total STAT3 levels were visualized by reprobing the membrane using a STAT3 antibody. Involvement of JAK in the dampening of acute <t>TNF-α</t> release by apoptotic cell supernatants. Monocytes were treated with apopt. SN for 3 h with or without a prior incubation with JAK inhibitor 1 for 15 min. Monocytes were then stimulated with LPS for 6 h and TNF-α levels in the culture media were measured by ELISA. STAT3 dependence of the inhibitory effect of apoptotic cell supernatants. Monocytes were pre-treated with the specific STAT3 Inhibitor VI for 30 min prior to treatment with the apoptotic neutrophil supernatant for 3 h and the LPS stimulation for 6 h. ELISA data are presented as the mean ± SEM of four independent experiments. The significance of the differences was evaluated by one-way ANOVA followed by Bonferroni's Multiple Comparison Test. p -values
    Lps Induced Acute Inflammation, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced acute inflammation/product/SLIT2 LTD
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lps induced acute inflammation - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    91
    Schiff Nutrition International lps induced aki mice periodicacid schiff pas staining
    Effect of 5b on lipopolysaccharide <t>(LPS)-induced</t> acute kidney injury <t>(AKI)</t> mice. <t>Periodicacid-Schiff</t> (PAS)-stained sections showed no damage in the vehicle group, the tubular injury in LPS group, and attenuated tubular injury in the 5b + LPS group. Note there were many dilated tubules (asterisk) in LPS-induced AKI mice. Kidney damage score was expressed as means ± SD for groups of six mice. *** p
    Lps Induced Aki Mice Periodicacid Schiff Pas Staining, supplied by Schiff Nutrition International, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced aki mice periodicacid schiff pas staining/product/Schiff Nutrition International
    Average 91 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    lps induced aki mice periodicacid schiff pas staining - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    99
    Millipore lps induced aki
    Effect of 5b on lipopolysaccharide <t>(LPS)-induced</t> acute kidney injury <t>(AKI)</t> mice. <t>Periodicacid-Schiff</t> (PAS)-stained sections showed no damage in the vehicle group, the tubular injury in LPS group, and attenuated tubular injury in the 5b + LPS group. Note there were many dilated tubules (asterisk) in LPS-induced AKI mice. Kidney damage score was expressed as means ± SD for groups of six mice. *** p
    Lps Induced Aki, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced aki/product/Millipore
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    lps induced aki - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    90
    Vital River Laboratories lps induced ali male balb c mice
    Network anti-inflammatory mechanism of JGT on <t>LPS-induced</t> <t>ALI.</t>
    Lps Induced Ali Male Balb C Mice, supplied by Vital River Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced ali male balb c mice/product/Vital River Laboratories
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    lps induced ali male balb c mice - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    89
    Orient Bio Company lps induced ali mouse model female balb c mice
    Effects of streptochlorin on <t>LPS-induced</t> <t>ALI</t> mouse model. <t>BALB/c</t> mice were given an intraperitoneal injection of streptochlorin successively 12 h and 2 h before being treated with LPS. ( A – D ) Histological examination of lung tissues was performed 24 h after the LPS challenge; Broncho-alveolar lavage fluid (BALF) was collected at 24 h after the LPS challenge to measure the total number of cells ( E ) and GR-1 + cells ( F ) were measured by flow cytometry; The concentrations of TNF-α ( G ) and IL-6 ( H ) in BALF after LPS prime alone or with indicated amounts of streptochlorin were measured by ELISA. The results are expressed as mean ± SEM. * p
    Lps Induced Ali Mouse Model Female Balb C Mice, supplied by Orient Bio Company, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lps induced ali mouse model female balb c mice/product/Orient Bio Company
    Average 89 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    lps induced ali mouse model female balb c mice - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    Image Search Results


    miR-147 is a negative regulator of macrophage inflammatory responses. ( A–C ) miR-147 mimics attenuate LPS, PAM3CSK4, and poly(I:C) induced inflammatory response in primary macrophages. Peritoneal macrophages were transfected with 40 nM control

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

    Article Title: miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses

    doi: 10.1073/pnas.0901216106

    Figure Lengend Snippet: miR-147 is a negative regulator of macrophage inflammatory responses. ( A–C ) miR-147 mimics attenuate LPS, PAM3CSK4, and poly(I:C) induced inflammatory response in primary macrophages. Peritoneal macrophages were transfected with 40 nM control

    Article Snippet: These data suggest that LPS-induced miR-147 expression requires de novo protein synthesis.

    Techniques: Transfection

    The miR-147 promoter is responsive to LPS stimulation. ( A ) Schematic representation of the miR-147 promoter, the locations of various transcriptional factor binding sites, the luciferase reporters containing the 2-kb miR-147 promoter and reporters with

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

    Article Title: miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses

    doi: 10.1073/pnas.0901216106

    Figure Lengend Snippet: The miR-147 promoter is responsive to LPS stimulation. ( A ) Schematic representation of the miR-147 promoter, the locations of various transcriptional factor binding sites, the luciferase reporters containing the 2-kb miR-147 promoter and reporters with

    Article Snippet: These data suggest that LPS-induced miR-147 expression requires de novo protein synthesis.

    Techniques: Binding Assay, Luciferase

    miR-147 is up-regulated in LPS-treated macrophages. ( A and B ) Time course of mature ( A ) and primary miR-147 ( B ) induction after LPS stimulation. Peritoneal macrophages were treated with 1 μg/ml LPS for the indicated lengths of time. Real-time

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

    Article Title: miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses

    doi: 10.1073/pnas.0901216106

    Figure Lengend Snippet: miR-147 is up-regulated in LPS-treated macrophages. ( A and B ) Time course of mature ( A ) and primary miR-147 ( B ) induction after LPS stimulation. Peritoneal macrophages were treated with 1 μg/ml LPS for the indicated lengths of time. Real-time

    Article Snippet: These data suggest that LPS-induced miR-147 expression requires de novo protein synthesis.

    Techniques:

    miR-147 is induced by stimulation of multiple TLRs. ( A ) Dose-dependent induction of miR-147 by poly(I:C) and LPS. Peritoneal macrophages were treated with PAM3CSK4, poly(I:C) or LPS at the indicated concentrations for 6 h (PAM3CSK4 and LPS) or 9 h (poly(I:C)).

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

    Article Title: miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses

    doi: 10.1073/pnas.0901216106

    Figure Lengend Snippet: miR-147 is induced by stimulation of multiple TLRs. ( A ) Dose-dependent induction of miR-147 by poly(I:C) and LPS. Peritoneal macrophages were treated with PAM3CSK4, poly(I:C) or LPS at the indicated concentrations for 6 h (PAM3CSK4 and LPS) or 9 h (poly(I:C)).

    Article Snippet: These data suggest that LPS-induced miR-147 expression requires de novo protein synthesis.

    Techniques:

    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

    Article Snippet: To examine the role of Slit2 in regulating LPS-induced endothelial inflammation, we first analyzed its role in pro-inflammatory cytokine/chemokine expression.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection, Western Blot

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: To examine the role of Slit2 in regulating LPS-induced endothelial inflammation, we first analyzed its role in pro-inflammatory cytokine/chemokine expression.

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

    Article Snippet: To examine the role of Slit2 in regulating LPS-induced endothelial inflammation, we first analyzed its role in pro-inflammatory cytokine/chemokine expression.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling

    Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

    Article Snippet: To examine the role of Slit2 in regulating LPS-induced endothelial inflammation, we first analyzed its role in pro-inflammatory cytokine/chemokine expression.

    Techniques: Labeling

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: To examine the role of Slit2 in regulating LPS-induced endothelial inflammation, we first analyzed its role in pro-inflammatory cytokine/chemokine expression.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

    Article Snippet: Here we show that, in primary Human Umbilical Vein Endothelial Cells (HUVECs), Slit2 represses lipopolysaccharide (LPS)-induced secretion of certain inflammatory cytokines/chemokines, cell adhesion molecule ICAM-1 upregulation and monocyte adhesion.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection, Western Blot

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: Here we show that, in primary Human Umbilical Vein Endothelial Cells (HUVECs), Slit2 represses lipopolysaccharide (LPS)-induced secretion of certain inflammatory cytokines/chemokines, cell adhesion molecule ICAM-1 upregulation and monocyte adhesion.

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

    Article Snippet: Here we show that, in primary Human Umbilical Vein Endothelial Cells (HUVECs), Slit2 represses lipopolysaccharide (LPS)-induced secretion of certain inflammatory cytokines/chemokines, cell adhesion molecule ICAM-1 upregulation and monocyte adhesion.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling

    Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

    Article Snippet: Here we show that, in primary Human Umbilical Vein Endothelial Cells (HUVECs), Slit2 represses lipopolysaccharide (LPS)-induced secretion of certain inflammatory cytokines/chemokines, cell adhesion molecule ICAM-1 upregulation and monocyte adhesion.

    Techniques: Labeling

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: Here we show that, in primary Human Umbilical Vein Endothelial Cells (HUVECs), Slit2 represses lipopolysaccharide (LPS)-induced secretion of certain inflammatory cytokines/chemokines, cell adhesion molecule ICAM-1 upregulation and monocyte adhesion.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: Along with these pro-inflammatory cytokines, some LPS-induced anti-inflammatory cytokines (including sICAM-1 and IL-1Ra) were also repressed by Slit2 (data not shown).

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: Along with these pro-inflammatory cytokines, some LPS-induced anti-inflammatory cytokines (including sICAM-1 and IL-1Ra) were also repressed by Slit2 (data not shown).

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

    Article Snippet: Thus we set out to identify which Robo receptor is responsible for the effect of Slit2 on LPS-induced inflammation in endothelial cells.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection, Western Blot

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: Thus we set out to identify which Robo receptor is responsible for the effect of Slit2 on LPS-induced inflammation in endothelial cells.

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

    Article Snippet: Thus we set out to identify which Robo receptor is responsible for the effect of Slit2 on LPS-induced inflammation in endothelial cells.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling

    Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

    Article Snippet: Thus we set out to identify which Robo receptor is responsible for the effect of Slit2 on LPS-induced inflammation in endothelial cells.

    Techniques: Labeling

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: Thus we set out to identify which Robo receptor is responsible for the effect of Slit2 on LPS-induced inflammation in endothelial cells.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    Effects of lipopolysaccharide (LPS) on Slit2 expression. ( A ) Lung inflammation induced by the LPS treatment. ( B ) Expression of the Slit2-exon15 isoforms in the lungs and brains of mice treated with an intranasal administration of LPS. ( C ) Total expression level of Slit2 in the lungs relative to 18S rRNA. ( D ) Total expression level of Slit2 in the brain relative to 18S rRNA. (ΔCT = CT slit2 − CT 18SrRNA ; CT: cycle threshold). Higher ΔCT means a lower expression of Slit2. * p

    Journal: Cancers

    Article Title: Lung Tumorigenesis Alters the Expression of Slit2-exon15 Splicing Variants in Tumor Microenvironment

    doi: 10.3390/cancers11020166

    Figure Lengend Snippet: Effects of lipopolysaccharide (LPS) on Slit2 expression. ( A ) Lung inflammation induced by the LPS treatment. ( B ) Expression of the Slit2-exon15 isoforms in the lungs and brains of mice treated with an intranasal administration of LPS. ( C ) Total expression level of Slit2 in the lungs relative to 18S rRNA. ( D ) Total expression level of Slit2 in the brain relative to 18S rRNA. (ΔCT = CT slit2 − CT 18SrRNA ; CT: cycle threshold). Higher ΔCT means a lower expression of Slit2. * p

    Article Snippet: However, an injection of lung cancer cells via the tail vein and the LPS-induced lung inflammation both decreased the Slit2 expression.

    Techniques: Expressing, Mouse Assay

    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

    Article Snippet: Furthermore, as shown in , our analysis of the microarray data ( ) revealed that i.p. injection of LPS-induced significant downregulated Slit2 and Robo4 in the whole heart of mice, and that pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) treatment induced significant downregulation of Slit2 and Robo4 in HUVEC.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection, Western Blot

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: Furthermore, as shown in , our analysis of the microarray data ( ) revealed that i.p. injection of LPS-induced significant downregulated Slit2 and Robo4 in the whole heart of mice, and that pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) treatment induced significant downregulation of Slit2 and Robo4 in HUVEC.

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The modulation of Slit2, Robo1 and Robo4 expression and the role of miR-218 (A, B) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). qRT-PCR was used to detect mRNA expression of Slit2 and Robo4. In accordance with mRNA level, protein levels of Robo4 in treated HUVECs for 12 h were shown as WB. (C) Protein levels of Robo1 in the above treated HUVECs or Robo1 of HUVECs treated only with Slit2-N for 12 h were shown by WB. (D) miR-218 level was detected by miR-218-specific qRT-PCR primers in HUVECs treated with LPS in the presence or absence of Slit2-N (30 nmol/L), and that of HUVECs treated with Slit2-N or PBS as control for 2 h. (E) Slit2-N treatment per se induced Slit2 and miR-218 upregulation as shown by qRT-PCR. (F) Slit2-N treatment per se reduced Robo1 expression, without affecting Robo4. (G) Slit2 and Robo1 expression levels in 10 different human tissues were detected using qRT-PCR with human multiple tissue cDNA panel (ovary, prostate, kidney, colon, heart, small intestine, liver, pancreas, lung and skeletal muscle). Values were normalized to human normal tissue control. Each dot stands for one human tissue with Slit2 and Robo1 expression levels on two axes. Correlation coefficient rho and p values are labeled in the plot. (* p

    Article Snippet: Furthermore, as shown in , our analysis of the microarray data ( ) revealed that i.p. injection of LPS-induced significant downregulated Slit2 and Robo4 in the whole heart of mice, and that pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) treatment induced significant downregulation of Slit2 and Robo4 in HUVEC.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling

    Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

    Article Snippet: Furthermore, as shown in , our analysis of the microarray data ( ) revealed that i.p. injection of LPS-induced significant downregulated Slit2 and Robo4 in the whole heart of mice, and that pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) treatment induced significant downregulation of Slit2 and Robo4 in HUVEC.

    Techniques: Labeling

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: Furthermore, as shown in , our analysis of the microarray data ( ) revealed that i.p. injection of LPS-induced significant downregulated Slit2 and Robo4 in the whole heart of mice, and that pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) treatment induced significant downregulation of Slit2 and Robo4 in HUVEC.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2 inhibits LPS-induced Pyk2 activation and NF-kB activation and translocation (A) Surface levels of TLR4 on HUVECs with Slit2-N (30 nmol/L) for different time (0, 0.5, 1 and 3 h) were analyzed by flow cytometry. IgG isotype control is shown as a solid peak in the histogram, TLR4 levels at different time points as open peaks of colored lines. (B) Flow cytometry data is quantified by mean fluorescence intensity and summarized in 3B. Changes are not statistically significant. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Pyk2 inhibitor Tyrphostin A9 (Tyr A9, 10 μmol/L). Cell culture supernatants were collected after treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are relative net optical signal intensity and are not comparable between cytokines. (D) HUVECs were treated with 100 ng/mL LPS for 2 h with or without pre-treatment of Slit2-N (30 nmol/L). Phosphorylated Pyk2 was detected by WB using p-Pyk2 (Y402) and Pyk2 antibodies. GAPDH was used as loading control. (E) HUVECs were pre-treated with Pyk2 inhibitor PF431396 (PF, 10 μmol/L), the indicated concentration of TLR4 neutralizing antibody (TLR4.Neu.Ab) or their controls for 1h and then stimulated with LPS for 2 h. Protein expression was analyzed by qRT-PCR. (F) Representative cytoplasmic (CY) and nuclear (NU) portions of HUVEC cell lysates were probed for nuclear marker Oct1 (exclusively in nuclear portion) and cytoplasmic marker GAPDH (mostly in cytoplasmic portion). HUVECs were treated with 100 ng/mL LPS for 4 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were then lysed, nuclear and cytoplasmic portions were separated. Active NF-kB was detected by an ELISA-based assay. (* p

    Article Snippet: This indicates that Slit2 treatment inhibited LPS-induced Pyk2 activation and further decreased the downstream activation and nuclear translocation of NF-kB in HUVECs.

    Techniques: Activation Assay, Translocation Assay, Flow Cytometry, Fluorescence, Cell Culture, Western Blot, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Enzyme-linked Immunosorbent Assay

    Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Proposed mechanism of Slit2 regulating LPS-induced endothelial inflammation and dysregulation of Slit2-Robo pathway by LPS Pro-inflammatory TLR4-Pyk2-NF-kB and Slit2-Robo1 pathways are labeled as red shapes, anti-inflammatory Slit2-Robo4 as green. Robo4 is dominantly expressed in endothelial cells, compared to Robo1.

    Article Snippet: This indicates that Slit2 treatment inhibited LPS-induced Pyk2 activation and further decreased the downstream activation and nuclear translocation of NF-kB in HUVECs.

    Techniques: Labeling

    The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: The anti-inflammatory effect of Slit2 was dependent on Robo4 rather than Robo1 (A) Robo1 and Robo4 relative mRNA expression level in resting HUVECs were quantified by qRT-PCR, and PCR product was shown on Agarose Gel. (B) 48 h after transfection with Robo1-or Robo4-siRNA, HUVECs were lysed and WB was used to detect the protein levels of Robo1 and Robo4. GAPDH was used as loading control. (C and D) 48 h after Robo1 and Robo4 knocking down respectively, GM-CSF mRNA expression in HUVECs was analyzed by qRT-PCR 2 h after LPS (100 ng/mL) treatment with or without Slit2-N (30 nmol/L). Non-targeting siRNA was used as control for Robo1- or Robo4-siRNA. (* p

    Article Snippet: In contrast, Slit2 enhanced LPS-induced GM-CSF expression when Robo4 was knocked down in HUVECs , which showed that the anti-inflammatory effect of Slit2 was mediated through Robo4 receptor.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection, Western Blot

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

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

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: In contrast, Slit2 enhanced LPS-induced GM-CSF expression when Robo4 was knocked down in HUVECs , which showed that the anti-inflammatory effect of Slit2 was mediated through Robo4 receptor.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    The anti-inflammatory effects of apoptotic cells are mediated through activation of JAK/STAT3 signalling Apoptotic neutrophil culture supernatants (apopt. SN) suppress LPS-induced TNF-α release. Monocytes were pre-treated for 3 h with apopt. SN and subsequently stimulated with LPS for 6 h. TNF-α contents in the culture media were measured by TNF-α ELISA. Activation of STAT3. Monocytes were treated with culture supernatants from either fresh or apoptotic cells for 1 h. STAT3 activation was detected by immunoblotting of cell lysates with the anti-pTYR705-STAT3 antibody. Total STAT3 levels were visualized by reprobing the membrane using a STAT3 antibody. Involvement of JAK in the dampening of acute TNF-α release by apoptotic cell supernatants. Monocytes were treated with apopt. SN for 3 h with or without a prior incubation with JAK inhibitor 1 for 15 min. Monocytes were then stimulated with LPS for 6 h and TNF-α levels in the culture media were measured by ELISA. STAT3 dependence of the inhibitory effect of apoptotic cell supernatants. Monocytes were pre-treated with the specific STAT3 Inhibitor VI for 30 min prior to treatment with the apoptotic neutrophil supernatant for 3 h and the LPS stimulation for 6 h. ELISA data are presented as the mean ± SEM of four independent experiments. The significance of the differences was evaluated by one-way ANOVA followed by Bonferroni's Multiple Comparison Test. p -values

    Journal: EMBO Molecular Medicine

    Article Title: Annexin A1 released from apoptotic cells acts through formyl peptide receptors to dampen inflammatory monocyte activation via JAK/STAT/SOCS signalling

    doi: 10.1002/emmm.201000113

    Figure Lengend Snippet: The anti-inflammatory effects of apoptotic cells are mediated through activation of JAK/STAT3 signalling Apoptotic neutrophil culture supernatants (apopt. SN) suppress LPS-induced TNF-α release. Monocytes were pre-treated for 3 h with apopt. SN and subsequently stimulated with LPS for 6 h. TNF-α contents in the culture media were measured by TNF-α ELISA. Activation of STAT3. Monocytes were treated with culture supernatants from either fresh or apoptotic cells for 1 h. STAT3 activation was detected by immunoblotting of cell lysates with the anti-pTYR705-STAT3 antibody. Total STAT3 levels were visualized by reprobing the membrane using a STAT3 antibody. Involvement of JAK in the dampening of acute TNF-α release by apoptotic cell supernatants. Monocytes were treated with apopt. SN for 3 h with or without a prior incubation with JAK inhibitor 1 for 15 min. Monocytes were then stimulated with LPS for 6 h and TNF-α levels in the culture media were measured by ELISA. STAT3 dependence of the inhibitory effect of apoptotic cell supernatants. Monocytes were pre-treated with the specific STAT3 Inhibitor VI for 30 min prior to treatment with the apoptotic neutrophil supernatant for 3 h and the LPS stimulation for 6 h. ELISA data are presented as the mean ± SEM of four independent experiments. The significance of the differences was evaluated by one-way ANOVA followed by Bonferroni's Multiple Comparison Test. p -values

    Article Snippet: Anti-inflammatory mediators released from apoptotic cells activate the JAK/STAT pathway We first examined the effects of mediators released by apoptotic human neutrophils on the lipopolysaccharide (LPS, Calbiochem)-induced TNF-α release in monocytes.

    Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Incubation

    Effect of 5b on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) mice. Periodicacid-Schiff (PAS)-stained sections showed no damage in the vehicle group, the tubular injury in LPS group, and attenuated tubular injury in the 5b + LPS group. Note there were many dilated tubules (asterisk) in LPS-induced AKI mice. Kidney damage score was expressed as means ± SD for groups of six mice. *** p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Anti-Inflammatory Pyranochalcone Derivative Attenuates LPS-Induced Acute Kidney Injury via Inhibiting TLR4/NF-κB Pathway

    doi: 10.3390/molecules22101683

    Figure Lengend Snippet: Effect of 5b on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) mice. Periodicacid-Schiff (PAS)-stained sections showed no damage in the vehicle group, the tubular injury in LPS group, and attenuated tubular injury in the 5b + LPS group. Note there were many dilated tubules (asterisk) in LPS-induced AKI mice. Kidney damage score was expressed as means ± SD for groups of six mice. *** p

    Article Snippet: Effect of 5b on LPS-Induced AKI Mice Periodicacid-Schiff (PAS) staining showed normal kidney tubules in the vehicle group.

    Techniques: Mouse Assay, Staining

    Network anti-inflammatory mechanism of JGT on LPS-induced ALI.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Chemomics-Integrated Proteomics Analysis of Jie-Geng-Tang to Ameliorate Lipopolysaccharide-Induced Acute Lung Injury in Mice

    doi: 10.1155/2016/7379146

    Figure Lengend Snippet: Network anti-inflammatory mechanism of JGT on LPS-induced ALI.

    Article Snippet: Experimental Animals and LPS-Induced ALI Male BALB/c mice weighing 18–22 g were purchased from Vital River Company (Beijing, China) and housed in a unidirectional airflow room under controlled temperature (20–24°C), relative humidity (40–60%), and a 12 h light/dark cycle.

    Techniques:

    Effects of streptochlorin on LPS-induced ALI mouse model. BALB/c mice were given an intraperitoneal injection of streptochlorin successively 12 h and 2 h before being treated with LPS. ( A – D ) Histological examination of lung tissues was performed 24 h after the LPS challenge; Broncho-alveolar lavage fluid (BALF) was collected at 24 h after the LPS challenge to measure the total number of cells ( E ) and GR-1 + cells ( F ) were measured by flow cytometry; The concentrations of TNF-α ( G ) and IL-6 ( H ) in BALF after LPS prime alone or with indicated amounts of streptochlorin were measured by ELISA. The results are expressed as mean ± SEM. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Anti-Inflammatory Effect of Streptochlorin via TRIF-Dependent Signaling Pathways in Cellular and Mouse Models

    doi: 10.3390/ijms16046902

    Figure Lengend Snippet: Effects of streptochlorin on LPS-induced ALI mouse model. BALB/c mice were given an intraperitoneal injection of streptochlorin successively 12 h and 2 h before being treated with LPS. ( A – D ) Histological examination of lung tissues was performed 24 h after the LPS challenge; Broncho-alveolar lavage fluid (BALF) was collected at 24 h after the LPS challenge to measure the total number of cells ( E ) and GR-1 + cells ( F ) were measured by flow cytometry; The concentrations of TNF-α ( G ) and IL-6 ( H ) in BALF after LPS prime alone or with indicated amounts of streptochlorin were measured by ELISA. The results are expressed as mean ± SEM. * p

    Article Snippet: Animals and LPS-Induced ALI Mouse Model Female BALB/c mice (22–25 g, 8 weeks old) were purchased from Orient Bio (Seoul, Korea).

    Techniques: Mouse Assay, Injection, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay