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Pharmacodynamics of isoflurane in patients with hepatic carcinoma during the perioperative period. Expression levels of (A) <t>TNF-α</t> and (B) IL-2 in patients that underwent tumor resection following pretreatment with isoflurane. (C) Serum concentration of isoflurane in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. (D) Cmax concentrations of isoflurane (0–0.40 mg/kg) in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
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Images

1) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Pharmacodynamics of isoflurane in patients with hepatic carcinoma during the perioperative period. Expression levels of (A) TNF-α and (B) IL-2 in patients that underwent tumor resection following pretreatment with isoflurane. (C) Serum concentration of isoflurane in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. (D) Cmax concentrations of isoflurane (0–0.40 mg/kg) in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Pharmacodynamics of isoflurane in patients with hepatic carcinoma during the perioperative period. Expression levels of (A) TNF-α and (B) IL-2 in patients that underwent tumor resection following pretreatment with isoflurane. (C) Serum concentration of isoflurane in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. (D) Cmax concentrations of isoflurane (0–0.40 mg/kg) in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing, Concentration Assay

2) Product Images from "Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene"

Article Title: Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2018.12.016

Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .
Figure Legend Snippet: Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .

Techniques Used: Activation Assay, Transduction, Translocation Assay, Expressing, Permeability, In Vitro, In Vivo

Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P
Figure Legend Snippet: Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P

Techniques Used: Permeability, Activation Assay, Transfection, Expressing

Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P
Figure Legend Snippet: Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P

Techniques Used: Activation Assay, Transfection, Expressing, Immunofluorescence, Translocation Assay

3) Product Images from "Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene"

Article Title: Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2018.12.016

Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P
Figure Legend Snippet: Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P

Techniques Used: Activation Assay, Transfection, Expressing, Immunofluorescence, Translocation Assay

Role of transforming growth factor-β–activating kinase (TAK)-1 in the lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway and Caco-2 tight junction permeability. A: LPS treatment at the concentration of 300 pg/mL caused activation of phospho-TAK-1. B and C: LPS treatment did not induce phosphorylation of NF-κB–inducing kinase (NIK) or mitogen-activated kinase kinase (MEKK)-1 at day 3 after LPS exposure compared with untreated Caco-2 cells. D: Confocal immunofluorescence of Caco-2 cells treated with 300 pg/mL LPS for 5 days indicated increase in pTAK-1 (green) on day 3 and 3.5 after LPS treatment. Nucleus, blue. E: TAK-1, NIK, and MEKK-1 siRNA transfections in Caco-2 cells significantly reduced TAK-1, NIK, and MEKK-1 protein expression, as analyzed by Western blot analysis and relative densitometry, respectively. F: TAK-1 siRNA transfection prevented the LPS-induced drop in Caco-2 cell transepithelial electrical resistance (TER). G: TAK-1 siRNA transfection inhibited the LPS-induced increase in Caco-2 inulin flux. H: NIK or MEKK-1 siRNA in transfection in Caco-2 cells did not prevent the LPS-induced drop in Caco-2 TER. Data are expressed as means ± SEM. n = 4 experiments ( A ). ∗∗∗ P
Figure Legend Snippet: Role of transforming growth factor-β–activating kinase (TAK)-1 in the lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway and Caco-2 tight junction permeability. A: LPS treatment at the concentration of 300 pg/mL caused activation of phospho-TAK-1. B and C: LPS treatment did not induce phosphorylation of NF-κB–inducing kinase (NIK) or mitogen-activated kinase kinase (MEKK)-1 at day 3 after LPS exposure compared with untreated Caco-2 cells. D: Confocal immunofluorescence of Caco-2 cells treated with 300 pg/mL LPS for 5 days indicated increase in pTAK-1 (green) on day 3 and 3.5 after LPS treatment. Nucleus, blue. E: TAK-1, NIK, and MEKK-1 siRNA transfections in Caco-2 cells significantly reduced TAK-1, NIK, and MEKK-1 protein expression, as analyzed by Western blot analysis and relative densitometry, respectively. F: TAK-1 siRNA transfection prevented the LPS-induced drop in Caco-2 cell transepithelial electrical resistance (TER). G: TAK-1 siRNA transfection inhibited the LPS-induced increase in Caco-2 inulin flux. H: NIK or MEKK-1 siRNA in transfection in Caco-2 cells did not prevent the LPS-induced drop in Caco-2 TER. Data are expressed as means ± SEM. n = 4 experiments ( A ). ∗∗∗ P

Techniques Used: Activation Assay, Permeability, Concentration Assay, Immunofluorescence, Transfection, Expressing, Western Blot

4) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Pharmacodynamics of isoflurane in patients with hepatic carcinoma during the perioperative period. Expression levels of (A) TNF-α and (B) IL-2 in patients that underwent tumor resection following pretreatment with isoflurane. (C) Serum concentration of isoflurane in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. (D) Cmax concentrations of isoflurane (0–0.40 mg/kg) in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Pharmacodynamics of isoflurane in patients with hepatic carcinoma during the perioperative period. Expression levels of (A) TNF-α and (B) IL-2 in patients that underwent tumor resection following pretreatment with isoflurane. (C) Serum concentration of isoflurane in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. (D) Cmax concentrations of isoflurane (0–0.40 mg/kg) in patients with hepatic carcinoma that underwent tumor resection following pretreatment with isoflurane. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing, Concentration Assay

5) Product Images from "Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene"

Article Title: Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2018.12.016

Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .
Figure Legend Snippet: Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .

Techniques Used: Activation Assay, Transduction, Translocation Assay, Expressing, Permeability, In Vitro, In Vivo

Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P
Figure Legend Snippet: Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P

Techniques Used: Permeability, Activation Assay, Transfection, Expressing

Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P
Figure Legend Snippet: Effect of toll-like receptor (TLR)-4, myeloid differentiation primary response (MyD)88, and transforming growth factor-β–activating kinase (TAK)-1 siRNA on lipopolysaccharide (LPS)-induced activation of NF-κB canonical pathway in Caco-2 cells. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of inhibitory κ B (IκB)-α protein expression compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IκB-α protein levels. B: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced increase in TAK-1 phosphorylation. Relative densitometry analysis of pTAK-1 protein levels. C: Confocal immunofluorescence showed that TLR-4, MyD88, and TAK-1 siRNA transfection of Caco-2 monolayers prevented the LPS-induced p65 (red) translocation to the nucleus (blue) ( arrowhead ) at 3 day after LPS exposure D: TLR-4 siRNA and MyD88 siRNA transfection in Caco-2 cells prevented LPS-induced degradation of IκB-α protein expression. Densitometry of IκB-α protein levels. E: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced activation of IκB kinase (IKK)-α and IKK-β compared with nontarget (NT) siRNA-transfected LPS-treated cells. Relative densitometry analysis of IKK-α and IKK-β is also shown. n = 4 experiments. ∗∗ P

Techniques Used: Activation Assay, Transfection, Expressing, Immunofluorescence, Translocation Assay

6) Product Images from "Experimental chronic kidney disease attenuates ischemia-reperfusion injury in an ex vivo rat lung model"

Article Title: Experimental chronic kidney disease attenuates ischemia-reperfusion injury in an ex vivo rat lung model

Journal: PLoS ONE

doi: 10.1371/journal.pone.0171736

Effects of HSP70 on the protective effects of CKD against IR lung injury. (A) TNF-α level in the BAL; (B) TNF-α level in the perfusate; (C) IL-10 level in the perfusate; (D) HSP70 level in the lung tissue; (E) cytoplasmic HSP70 level in the lung tissue determined by western blotting; and (F) HSP70 level in the lung tissue determined by western blotting in IR induced lung injury. The data are expressed as the means ± SD. *Significantly different from the control ( P
Figure Legend Snippet: Effects of HSP70 on the protective effects of CKD against IR lung injury. (A) TNF-α level in the BAL; (B) TNF-α level in the perfusate; (C) IL-10 level in the perfusate; (D) HSP70 level in the lung tissue; (E) cytoplasmic HSP70 level in the lung tissue determined by western blotting; and (F) HSP70 level in the lung tissue determined by western blotting in IR induced lung injury. The data are expressed as the means ± SD. *Significantly different from the control ( P

Techniques Used: Western Blot

7) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Activity Assay, Expressing, Migration

8) Product Images from "Cellular fibronectin containing extra domain A promotes arterial thrombosis in mice through platelet Toll-like receptor 4"

Article Title: Cellular fibronectin containing extra domain A promotes arterial thrombosis in mice through platelet Toll-like receptor 4

Journal: Blood

doi: 10.1182/blood-2014-10-608653

Cellular Fn-EDA+ promotes carotid artery thrombosis through TLR4. Graphs representing mean time to first thrombus formation (A), thrombus growth (B), and mean time to occlusion (C) in FeCl 3 -injured carotid arteries. * P
Figure Legend Snippet: Cellular Fn-EDA+ promotes carotid artery thrombosis through TLR4. Graphs representing mean time to first thrombus formation (A), thrombus growth (B), and mean time to occlusion (C) in FeCl 3 -injured carotid arteries. * P

Techniques Used:

Platelet TLR4 contributes to cellular Fn-EDA+-mediated accelerated thrombosis in injured arteries. (A) Schematic depicting the technique for generating chimeric mice with platelet-specific TLR4 deficiency. (B-D) Graphs representing mean time to first
Figure Legend Snippet: Platelet TLR4 contributes to cellular Fn-EDA+-mediated accelerated thrombosis in injured arteries. (A) Schematic depicting the technique for generating chimeric mice with platelet-specific TLR4 deficiency. (B-D) Graphs representing mean time to first

Techniques Used: Mouse Assay

TLR4 on cells of hematopoietic origin contributes to cellular Fn-EDA+-mediated accelerated thrombosis. (A) Real-time PCR analysis for TLR4 (normalized with glyceraldehyde-3-phosphate dehydrogenase) in genomic DNA from peripheral blood mononuclear cells
Figure Legend Snippet: TLR4 on cells of hematopoietic origin contributes to cellular Fn-EDA+-mediated accelerated thrombosis. (A) Real-time PCR analysis for TLR4 (normalized with glyceraldehyde-3-phosphate dehydrogenase) in genomic DNA from peripheral blood mononuclear cells

Techniques Used: Real-time Polymerase Chain Reaction

Cellular Fn-EDA+ potentiates platelet aggregation through TLR4. (A) Representative immunoblots showing interaction between cellular Fn-EDA+ and TLR4. Proteins in platelet lysates from Fn-EDA +/+ or Fn-EDA −/− mice were immunoprecipitated
Figure Legend Snippet: Cellular Fn-EDA+ potentiates platelet aggregation through TLR4. (A) Representative immunoblots showing interaction between cellular Fn-EDA+ and TLR4. Proteins in platelet lysates from Fn-EDA +/+ or Fn-EDA −/− mice were immunoprecipitated

Techniques Used: Western Blot, Mouse Assay, Immunoprecipitation

9) Product Images from "Transferrin-dependent crosstalk between the intestinal tract and commensal microbes contributes for immune tolerance"

Article Title: Transferrin-dependent crosstalk between the intestinal tract and commensal microbes contributes for immune tolerance

Journal: bioRxiv

doi: 10.1101/2020.03.02.972281

Transferrin inhibits TLR4 activation. THP-1 cells were stimulated in presence or absence of transferrin by LPS for 8 h. Total and phosphorylated TAK1, IKKα, IκBα, and p65 of the myD88-dependent pathway and TBK1 and IRF3 of the myD88-independent pathway in THP-1 cells were analyzed by Western blotting, respectively. Corresponding quantifications are shown on right (six panels). β-actin was used as loading control. Data represent means ± SD of five independent experiments, * p
Figure Legend Snippet: Transferrin inhibits TLR4 activation. THP-1 cells were stimulated in presence or absence of transferrin by LPS for 8 h. Total and phosphorylated TAK1, IKKα, IκBα, and p65 of the myD88-dependent pathway and TBK1 and IRF3 of the myD88-independent pathway in THP-1 cells were analyzed by Western blotting, respectively. Corresponding quantifications are shown on right (six panels). β-actin was used as loading control. Data represent means ± SD of five independent experiments, * p

Techniques Used: Activation Assay, Western Blot

10) Product Images from "Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene"

Article Title: Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2018.12.016

Effects of genetic knockdown of p65/p50 and p52/p100 on lipopolysaccharide (LPS)-induced increase in intestinal epithelial tight junction permeability. A: p65 siRNA transfection resulted in a near-complete depletion of p65. Relative densitometry analysis of p65 protein expression levels. B: The siRNA of p50 transfection resulted in a near-complete depletion of p50 protein expression. Relative densitometry of p50 protein expression levels. C: The p52 siRNA transfection resulted in a near-complete depletion of p52 as shown in densitometry analysis. D: p100 siRNA transfection resulted in a near-complete depletion of p100, as shown in densitometry analysis. E: p65 and p50 siRNA transfection in Caco-2 monolayers inhibited the LPS-induced drop in Caco-2 cell transepithelial electrical resistance (TER). F: p65 and p50 siRNA transfection inhibited the LPS-induced increase in inulin flux. G: p52 and p100 siRNA transfection in Caco-2 monolayers did not prevent the LPS-induced drop in TER. H: p52 and p100 siRNA transfection did not prevent the LPS-induced increase in inulin flux. The Western blot analysis was performed 72 hours after all siRNA transfections. n = 4 experiments ( A–D and F ); n = 3 experiments ( H ). ∗∗∗ P
Figure Legend Snippet: Effects of genetic knockdown of p65/p50 and p52/p100 on lipopolysaccharide (LPS)-induced increase in intestinal epithelial tight junction permeability. A: p65 siRNA transfection resulted in a near-complete depletion of p65. Relative densitometry analysis of p65 protein expression levels. B: The siRNA of p50 transfection resulted in a near-complete depletion of p50 protein expression. Relative densitometry of p50 protein expression levels. C: The p52 siRNA transfection resulted in a near-complete depletion of p52 as shown in densitometry analysis. D: p100 siRNA transfection resulted in a near-complete depletion of p100, as shown in densitometry analysis. E: p65 and p50 siRNA transfection in Caco-2 monolayers inhibited the LPS-induced drop in Caco-2 cell transepithelial electrical resistance (TER). F: p65 and p50 siRNA transfection inhibited the LPS-induced increase in inulin flux. G: p52 and p100 siRNA transfection in Caco-2 monolayers did not prevent the LPS-induced drop in TER. H: p52 and p100 siRNA transfection did not prevent the LPS-induced increase in inulin flux. The Western blot analysis was performed 72 hours after all siRNA transfections. n = 4 experiments ( A–D and F ); n = 3 experiments ( H ). ∗∗∗ P

Techniques Used: Permeability, Transfection, Expressing, Western Blot

Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .
Figure Legend Snippet: Proposed scheme of the intracellular pathways involved in lipopolysaccharide (LPS)-induced activation of canonical NF-κB pathway. LPS treatment results in activation of the toll-like receptor (TLR)-4 signal transduction and myeloid differentiation primary response (MyD)88-dependent signaling cascade. Activation of TLR-4/MyD88 signal transduction pathway leads to the activation of IL-1 receptor-associated kinase (IRAK)-4 and phosphorylation of transforming growth factor-β–activating kinase (TAK)-1, leading to the activation of canonical NF-κB pathway of both inhibitory κ B (IκB) kinases (IKKs), IKK-α and IKK-β, which further leads to degradation of IκB-α and nuclear translocation of NF-κB p65/p50. The activation of TAK-1 and canonical NF-κB p65/p50 causes up-regulation of myosin light chain kinase (MLCK) gene and protein expression, ultimately increasing tight junction (TJ) permeability in vitro and in vivo .

Techniques Used: Activation Assay, Transduction, Translocation Assay, Expressing, Permeability, In Vitro, In Vivo

Effect of genetic knockdown of (p65/p50) canonical pathway and transforming growth factor-β–activating kinase (TAK)-1 on lipopolysaccharide (LPS)-induced activation of myosin light chain kinase (MLCK) protein expression. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced increase in MLCK mRNA. B: Similarly, TAK-1 siRNA transfection in Caco-2 cells also inhibited LPS-induced increase in MLCK and phosphoMLC protein expression by Western blot analysis and was shown by relative densitometry of MLCK protein. C: p65 siRNA transfection in Caco-2 cells prevented LPS-induced increase in MLCK mRNA. D: p50 and p65 siRNA transfection in Caco-2 cells caused marked inhibition of LPS-induced increase in MLCK protein expression, as analyzed by Western blot analysis and relative analysis of MLCK protein levels by densitometry. n = 3 experiments ( A ); n = 4 experiments ( B and D ). ∗∗ P
Figure Legend Snippet: Effect of genetic knockdown of (p65/p50) canonical pathway and transforming growth factor-β–activating kinase (TAK)-1 on lipopolysaccharide (LPS)-induced activation of myosin light chain kinase (MLCK) protein expression. A: TAK-1 siRNA transfection in Caco-2 cells prevented LPS-induced increase in MLCK mRNA. B: Similarly, TAK-1 siRNA transfection in Caco-2 cells also inhibited LPS-induced increase in MLCK and phosphoMLC protein expression by Western blot analysis and was shown by relative densitometry of MLCK protein. C: p65 siRNA transfection in Caco-2 cells prevented LPS-induced increase in MLCK mRNA. D: p50 and p65 siRNA transfection in Caco-2 cells caused marked inhibition of LPS-induced increase in MLCK protein expression, as analyzed by Western blot analysis and relative analysis of MLCK protein levels by densitometry. n = 3 experiments ( A ); n = 4 experiments ( B and D ). ∗∗ P

Techniques Used: Activation Assay, Expressing, Transfection, Western Blot, Inhibition

Activation of p65/p50 canonical pathway by lipopolysaccharide (LPS) in mice enterocytes and effect of NF-κB inhibitors on LPS-induced increase in mouse intestinal epithelial tight junction permeability. A: LPS i.p. injections (0.1 mg/kg body weight) in mice caused activation of canonical p65/p50 pathway, as assessed by degradation of inhibitory κ B (IκB)-α protein expression on day 3 in mice enterocytes. Densitometry of IκB-α protein levels. B: The immunoblot analysis from LPS-treated mice enterocytes revealed significant increase in nuclear p65 protein expression on day 3 compared with untreated mice enterocytes. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Lamin B were used as loading controls for cytoplasmic (cyto) and nuclear (nuc) fractions, respectively C: Confocal immunofluorescence of mouse intestines treated with LPS (0.1 mg/kg body weight) on day 3 indicated p65 (red) ( arrowheads ) translocation to the nucleus (blue) compared with control (C) mouse enterocytes. D: NF-κB inhibitor, ammonium pyrrolidinedithiocarbamate (PDTC; 10 mg/kg body weight), and Bay-11 (5 mg/kg body weight) pretreatment prevented the LPS-induced increase in 10K dextran flux. PDTC and Bay-11 were dissolved in dimethyl sulfoxide and injected 1 hour before LPS treatment. Data are expressed as means ± SEM. n = 4 experiments ( A and C ); n = 3 experiments ( B ). ∗∗ P
Figure Legend Snippet: Activation of p65/p50 canonical pathway by lipopolysaccharide (LPS) in mice enterocytes and effect of NF-κB inhibitors on LPS-induced increase in mouse intestinal epithelial tight junction permeability. A: LPS i.p. injections (0.1 mg/kg body weight) in mice caused activation of canonical p65/p50 pathway, as assessed by degradation of inhibitory κ B (IκB)-α protein expression on day 3 in mice enterocytes. Densitometry of IκB-α protein levels. B: The immunoblot analysis from LPS-treated mice enterocytes revealed significant increase in nuclear p65 protein expression on day 3 compared with untreated mice enterocytes. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Lamin B were used as loading controls for cytoplasmic (cyto) and nuclear (nuc) fractions, respectively C: Confocal immunofluorescence of mouse intestines treated with LPS (0.1 mg/kg body weight) on day 3 indicated p65 (red) ( arrowheads ) translocation to the nucleus (blue) compared with control (C) mouse enterocytes. D: NF-κB inhibitor, ammonium pyrrolidinedithiocarbamate (PDTC; 10 mg/kg body weight), and Bay-11 (5 mg/kg body weight) pretreatment prevented the LPS-induced increase in 10K dextran flux. PDTC and Bay-11 were dissolved in dimethyl sulfoxide and injected 1 hour before LPS treatment. Data are expressed as means ± SEM. n = 4 experiments ( A and C ); n = 3 experiments ( B ). ∗∗ P

Techniques Used: Activation Assay, Mouse Assay, Permeability, Expressing, Immunofluorescence, Translocation Assay, Injection

11) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing

Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Activity Assay, Expressing, Migration

12) Product Images from "Renin-angiotensin system activation accelerates atherosclerosis in experimental renal failure by promoting endoplasmic reticulum stress-related inflammation"

Article Title: Renin-angiotensin system activation accelerates atherosclerosis in experimental renal failure by promoting endoplasmic reticulum stress-related inflammation

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2017.2856

Effects of inositol-requiring 1α (IRE1α) siRNA on the angiotensin II (Ang II)-induced inflammatory response in RAW264.7 macrophages. The RAW264.7 macrophages transfected with or without IRE1α siRNA were stimulated with 1 μ g/ml Ang II for 8 h and then subjected to western blot analysis and gene expression analysis. Western blot analysis of the expression of (A and B) IRE1α, p-IKK, IKKα/β, IκB, and (C and D) NF-κB p65. (E) RT-qPCR analysis of the gene expression of the indicated cytokines. The results were expressed as the mean ± SD. n=3 independent experiments; ** P
Figure Legend Snippet: Effects of inositol-requiring 1α (IRE1α) siRNA on the angiotensin II (Ang II)-induced inflammatory response in RAW264.7 macrophages. The RAW264.7 macrophages transfected with or without IRE1α siRNA were stimulated with 1 μ g/ml Ang II for 8 h and then subjected to western blot analysis and gene expression analysis. Western blot analysis of the expression of (A and B) IRE1α, p-IKK, IKKα/β, IκB, and (C and D) NF-κB p65. (E) RT-qPCR analysis of the gene expression of the indicated cytokines. The results were expressed as the mean ± SD. n=3 independent experiments; ** P

Techniques Used: Transfection, Western Blot, Expressing, Quantitative RT-PCR

Effects of losartan treatment on vascular inflammation and ER stress in uremic mice. The co-localization of CD68 and p-inositol-requiring 1α (IRE1α) was examined by immunohistochemistry using adjacent sections. (A and B) Quantitative analysis of macrophages content in lesion plaques. Immunohistochemistry with CD68, original magnification, ×40. n=10, 11, 10 for the control, SNx and SNx + losartan groups, respectively. (C and D) Quantitative analysis of p-IRE1α in lesion plaques in mice. Immunohistochemistry with p-IRE1α; original magnification, ×40. (E and F) Western blot analysis of the activation of IRE1α and the expression of glucose-regulated protein 78 (GRP78) in aortas of mice. n=5, 6, 5 for the control, SNx and SNx + losartan groups, respectively. (G) RT-qPCR analysis of pro-inflammatory cytokine and chemokine gene expression in aortas. n=5, 5, 5 for the control, SNx and SNx + losartan groups, respectively. Data are the means ± SEM. ** P
Figure Legend Snippet: Effects of losartan treatment on vascular inflammation and ER stress in uremic mice. The co-localization of CD68 and p-inositol-requiring 1α (IRE1α) was examined by immunohistochemistry using adjacent sections. (A and B) Quantitative analysis of macrophages content in lesion plaques. Immunohistochemistry with CD68, original magnification, ×40. n=10, 11, 10 for the control, SNx and SNx + losartan groups, respectively. (C and D) Quantitative analysis of p-IRE1α in lesion plaques in mice. Immunohistochemistry with p-IRE1α; original magnification, ×40. (E and F) Western blot analysis of the activation of IRE1α and the expression of glucose-regulated protein 78 (GRP78) in aortas of mice. n=5, 6, 5 for the control, SNx and SNx + losartan groups, respectively. (G) RT-qPCR analysis of pro-inflammatory cytokine and chemokine gene expression in aortas. n=5, 5, 5 for the control, SNx and SNx + losartan groups, respectively. Data are the means ± SEM. ** P

Techniques Used: Mouse Assay, Immunohistochemistry, Western Blot, Activation Assay, Expressing, Quantitative RT-PCR

Effects of losartan on the expression of ER stress marker proteins and nuclear factor-κB (NF-κB) inhibitor protein IκB in angiotensin II (Ang II)-stimulated RAW264.7 macrophages. RAW264.7 macrophages were stimulated with 1 μ g/ml Ang II for 4, 8, 16 and 24 h [for inositol-requiring 1α (IRE1α) activation (A and B); for glucose-regulated protein 78 (GRP78) (E and F)] or with 0.01, 0.1, 1, or 10 μ g/ml Ang II for 24 h [for IRE1α activation (C and D); for GRP78 (G and H)] and examined by western blot analysis. Culture medium was used as a blank control. (I–M) Western blot analysis of the activation of IRE1α, IKKα/β and IκB. The results were expressed as the means ± SD. n=3 independent experiments; * P
Figure Legend Snippet: Effects of losartan on the expression of ER stress marker proteins and nuclear factor-κB (NF-κB) inhibitor protein IκB in angiotensin II (Ang II)-stimulated RAW264.7 macrophages. RAW264.7 macrophages were stimulated with 1 μ g/ml Ang II for 4, 8, 16 and 24 h [for inositol-requiring 1α (IRE1α) activation (A and B); for glucose-regulated protein 78 (GRP78) (E and F)] or with 0.01, 0.1, 1, or 10 μ g/ml Ang II for 24 h [for IRE1α activation (C and D); for GRP78 (G and H)] and examined by western blot analysis. Culture medium was used as a blank control. (I–M) Western blot analysis of the activation of IRE1α, IKKα/β and IκB. The results were expressed as the means ± SD. n=3 independent experiments; * P

Techniques Used: Expressing, Marker, Activation Assay, Western Blot

13) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Activity Assay, Expressing, Migration

14) Product Images from "Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene"

Article Title: Lipopolysaccharide-Induced Increase in Intestinal Permeability Is Mediated by TAK-1 Activation of IKK and MLCK/MYLK Gene

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2018.12.016

Time course effect of lipopolysaccharide (LPS) on Caco-2 NF-κB pathway (p65 and p52) activation. A: LPS at the concentration of 300 pg/mL caused significant increase in p65 protein expression in Caco-2 cells on day 3 to 3.5. The expression of p52 and p100 did not change with LPS treatment. β-Actin was used as an internal control for protein loading. B: Relative densitometry analysis for p65 protein levels. C: Confocal immunofluorescence of Caco-2 cells treated with 300 pg/mL LPS (3 to 3.5 days) indicated p65 (red) translocation to the nucleus (blue) ( arrowheads ). D: P52 (green) did not change after LPS treatment ( arowheads ). E: LPS caused degradation of inhibitory κ B (IκB)-α expression (3 to 3.5 days), as assessed by Western blot analysis. β-Actin was used as an internal control for protein loading. F: Densitometry analysis of LPS treatment showed significant decrease in IκB-α level on day 3 to 3.5 compared with control untreated cells. G and H: LPS-treated Caco-2 cells (3 to 3.5 days) analyzed by flow cytometry for phospho-IκB-α and phospho-p65, respectively, showed increased expression compared with control untreated cells. Gray indicates isotope control (Cont); blue, control untreated; red, LPS treated day 3. [Mean fluorescence intensity (MFI): pIκB-α, 175 ± 12.12, versus 292 ± 25.63; pp65, 3307 ± 154.7, versus 4329.3 ± 169.4]. Data are expressed as means ± SEM. n = 3 independent experiments. ∗∗∗ P
Figure Legend Snippet: Time course effect of lipopolysaccharide (LPS) on Caco-2 NF-κB pathway (p65 and p52) activation. A: LPS at the concentration of 300 pg/mL caused significant increase in p65 protein expression in Caco-2 cells on day 3 to 3.5. The expression of p52 and p100 did not change with LPS treatment. β-Actin was used as an internal control for protein loading. B: Relative densitometry analysis for p65 protein levels. C: Confocal immunofluorescence of Caco-2 cells treated with 300 pg/mL LPS (3 to 3.5 days) indicated p65 (red) translocation to the nucleus (blue) ( arrowheads ). D: P52 (green) did not change after LPS treatment ( arowheads ). E: LPS caused degradation of inhibitory κ B (IκB)-α expression (3 to 3.5 days), as assessed by Western blot analysis. β-Actin was used as an internal control for protein loading. F: Densitometry analysis of LPS treatment showed significant decrease in IκB-α level on day 3 to 3.5 compared with control untreated cells. G and H: LPS-treated Caco-2 cells (3 to 3.5 days) analyzed by flow cytometry for phospho-IκB-α and phospho-p65, respectively, showed increased expression compared with control untreated cells. Gray indicates isotope control (Cont); blue, control untreated; red, LPS treated day 3. [Mean fluorescence intensity (MFI): pIκB-α, 175 ± 12.12, versus 292 ± 25.63; pp65, 3307 ± 154.7, versus 4329.3 ± 169.4]. Data are expressed as means ± SEM. n = 3 independent experiments. ∗∗∗ P

Techniques Used: Activation Assay, Concentration Assay, Expressing, Immunofluorescence, Translocation Assay, Western Blot, Flow Cytometry, Fluorescence

Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P
Figure Legend Snippet: Effect of lipopolysaccharide (LPS) on inhibitory κ B kinases (IKKs) and intestinal epithelial tight junction permeability. A: LPS at 300 pg/mL in Caco-2 cells caused significant increase in activation of both IKK-α and IKK-β on day 3 to 3.5 after LPS treatment. β-Actin was used as an internal control (C; Con; Cont) for protein loading. Densitometry analysis of LPS treatment showed significant increase in IKK-α and IKK-β, respectively, on day 3 and 3.5 compared with control untreated cells. B and C: siRNA transfection of IKK-α and IKK-β in Caco-2 cells inhibited the protein expression of IKK-α and IKK-β. D: siRNA transfection of IKK-α in Caco-2 cells partially prevented the LPS-induced increase in inulin flux. E: siRNA transfection of IKK-β in Caco-2 cells inhibited the LPS-induced increase in inulin flux. Data are expressed as means ± SEM. n = 6 experiments ( B , C , and E ). ∗∗∗ P

Techniques Used: Permeability, Activation Assay, Transfection, Expressing

15) Product Images from "Genistein suppresses tumor necrosis factor ?-induced inflammation via modulating reactive oxygen species/Akt/nuclear factor ?B and adenosine monophosphate-activated protein kinase signal pathways in human synoviocyte MH7A cells"

Article Title: Genistein suppresses tumor necrosis factor ?-induced inflammation via modulating reactive oxygen species/Akt/nuclear factor ?B and adenosine monophosphate-activated protein kinase signal pathways in human synoviocyte MH7A cells

Journal: Drug Design, Development and Therapy

doi: 10.2147/DDDT.S52354

( A  and  B ) Effects of genistein (Gen) on nuclear factor (NF)-κB activation in MH7A cells. Notes:  ( A ) MH7A cells were pretreated with Gen or not for 2 hours, and then incubated with tumor necrosis factor (TNF)-α for the indicated times. Phosphorylated IκB kinase (p-IKK)-α/β, p-IκBα, and p-NF-κB p65 levels were determined by Western blot analysis. ( B ) After being pretreated with Gen or not for 2 hours, MH7A cells were then incubated with TNF-α for 15 minutes. The nuclei translocation of NF-κB p65 was assessed by confocal fluorescence microscopy. Cells were immunostained with NF-κB p65 antibody. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments. Bar 10 μm. Abbreviation:  GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Figure Legend Snippet: ( A and B ) Effects of genistein (Gen) on nuclear factor (NF)-κB activation in MH7A cells. Notes: ( A ) MH7A cells were pretreated with Gen or not for 2 hours, and then incubated with tumor necrosis factor (TNF)-α for the indicated times. Phosphorylated IκB kinase (p-IKK)-α/β, p-IκBα, and p-NF-κB p65 levels were determined by Western blot analysis. ( B ) After being pretreated with Gen or not for 2 hours, MH7A cells were then incubated with TNF-α for 15 minutes. The nuclei translocation of NF-κB p65 was assessed by confocal fluorescence microscopy. Cells were immunostained with NF-κB p65 antibody. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments. Bar 10 μm. Abbreviation: GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Techniques Used: Activation Assay, Incubation, Western Blot, Translocation Assay, Fluorescence, Microscopy, Staining

16) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing

Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Activity Assay, Expressing, Migration

17) Product Images from "Impairment of hypoxia-induced angiogenesis by LDL involves a HIF-centered signaling network linking inflammatory TNFα and angiogenic VEGF"

Article Title: Impairment of hypoxia-induced angiogenesis by LDL involves a HIF-centered signaling network linking inflammatory TNFα and angiogenic VEGF

Journal: Aging (Albany NY)

doi: 10.18632/aging.101726

Knockdown of either HIF-1α or HIF-2α impairs hypoxia-induced angiogenesis and VEGF production in ECs. ( A ) HUVECs were transfected with pLKO.1 constructs encoding shRNA specifically targeting HIF-1α and HIF-2α, as well as scramble sequence as negative control (shNC). As the basal levels of HIF-1α or HIF-2α in HUVECs were relatively low, these transfected cells were exposed to 1% O 2 (21% O 2 as normoxia control) for 24 hrs, after which Western blot analysis was performed to confirm shRNA knockdown of HIF-1α and HIF-2α, respectively. ( B , C ) HUVECs expressing HIF-1α or HIF-2α shRNA were then exposed to 1% O 2 for the indicated intervals, followed by colony formation assay ( B , 72 hrs) and Matrigel-based tube formation assay ( C , 24 hrs). Representative microscopic images for at least three independent experiments were shown. Arrowheads indicate unclosed loops of vascular structure. ( D ) In parallel, the VEGF level was measured by ELISA assay after cultured for 72 hrs under 1% O 2 . Values represent the means ± SD for at least three independent experiments performed in triplicate. ** P
Figure Legend Snippet: Knockdown of either HIF-1α or HIF-2α impairs hypoxia-induced angiogenesis and VEGF production in ECs. ( A ) HUVECs were transfected with pLKO.1 constructs encoding shRNA specifically targeting HIF-1α and HIF-2α, as well as scramble sequence as negative control (shNC). As the basal levels of HIF-1α or HIF-2α in HUVECs were relatively low, these transfected cells were exposed to 1% O 2 (21% O 2 as normoxia control) for 24 hrs, after which Western blot analysis was performed to confirm shRNA knockdown of HIF-1α and HIF-2α, respectively. ( B , C ) HUVECs expressing HIF-1α or HIF-2α shRNA were then exposed to 1% O 2 for the indicated intervals, followed by colony formation assay ( B , 72 hrs) and Matrigel-based tube formation assay ( C , 24 hrs). Representative microscopic images for at least three independent experiments were shown. Arrowheads indicate unclosed loops of vascular structure. ( D ) In parallel, the VEGF level was measured by ELISA assay after cultured for 72 hrs under 1% O 2 . Values represent the means ± SD for at least three independent experiments performed in triplicate. ** P

Techniques Used: Transfection, Construct, shRNA, Sequencing, Negative Control, Western Blot, Expressing, Colony Assay, Tube Formation Assay, Enzyme-linked Immunosorbent Assay, Cell Culture

Native LDL, but not oxLDL, inhibits activation of HIF and NF−κB signals induced by hypoxia. HUVECs were treated as follows: ( A ) pre-treated with LDL (100 μg/ml) for 48 hrs, followed by incubation under hypoxic (1% O 2 ) condition for additional 48 hrs; ( B ) pre-treated with LDL (100 μg/ml) in the presence or absence of the free radical scavenger PBN (2 mM) for 48 hrs, and then exposed to the PHD inhibitor DMOG (1 μM) for additional 72 hrs; ( C ) pre-treated with the indicated concentrations of oxidized LDL (oxLDL, μg/ml) for 48 hrs, followed by DMOG (1 μM) for additional 72 hrs. After treatment, Western blot analysis was performed to monitor expression of HIF-1α, HIF-2α, and HIF-1β, as well as phosphorylation of NF-κB p65 (S536). ( D ) Alternatively, HUVECs were treated as described in panel 4B, after which cytoplasmic and nuclear fractions were separated and subjected to Western blot analysis for monitoring nuclear translocation of HIF-1α, HIF-2α, and HIF-1β. Blots were reprobed for β-actin and laminin A as loading controls for cytoplasmic and nuclear fractions, respectively. ( E ) HUVECs were pre-incubated with the NF-κB inhibitor PDTC (100 μM) for 4 hrs, followed by DMOG (1 μM) for additional 72 hrs, after which the protein levels of HIFs were monitored by Western blot analysis.
Figure Legend Snippet: Native LDL, but not oxLDL, inhibits activation of HIF and NF−κB signals induced by hypoxia. HUVECs were treated as follows: ( A ) pre-treated with LDL (100 μg/ml) for 48 hrs, followed by incubation under hypoxic (1% O 2 ) condition for additional 48 hrs; ( B ) pre-treated with LDL (100 μg/ml) in the presence or absence of the free radical scavenger PBN (2 mM) for 48 hrs, and then exposed to the PHD inhibitor DMOG (1 μM) for additional 72 hrs; ( C ) pre-treated with the indicated concentrations of oxidized LDL (oxLDL, μg/ml) for 48 hrs, followed by DMOG (1 μM) for additional 72 hrs. After treatment, Western blot analysis was performed to monitor expression of HIF-1α, HIF-2α, and HIF-1β, as well as phosphorylation of NF-κB p65 (S536). ( D ) Alternatively, HUVECs were treated as described in panel 4B, after which cytoplasmic and nuclear fractions were separated and subjected to Western blot analysis for monitoring nuclear translocation of HIF-1α, HIF-2α, and HIF-1β. Blots were reprobed for β-actin and laminin A as loading controls for cytoplasmic and nuclear fractions, respectively. ( E ) HUVECs were pre-incubated with the NF-κB inhibitor PDTC (100 μM) for 4 hrs, followed by DMOG (1 μM) for additional 72 hrs, after which the protein levels of HIFs were monitored by Western blot analysis.

Techniques Used: Activation Assay, Incubation, Western Blot, Expressing, Translocation Assay

The GEP analyses map the potential correlations among the HIFs, TNFα, VEGFs, and NF-κB signaling pathways in ECs. ( A , B ) By analyzing the datasets involving gene expression profiling (GEP) in ECs using the R2: Genomics Analysis and Visualization Platform, the heatmaps were generated based on P values for analyses of correlations (red and blue indicating positive and negative correlations, respectively) between each pair of genes as indicated on longitudinal and transverse axes, in ( A ; dataset, Exp HUVEC vs ESC - James - 12 - MAS5.0 - u133p2) human stem cells (hESC) vs human umbilical vein endothelial cells (HUVEC) and ( B ; dataset, Normal Endothelial Cells HUAEC/HUVEC - Luttun - 38 - MAS5.0 - u133p2) HUVEC vs human umbilical artery endothelial cells (HUAEC). Numbers in the heatmaps indicate the areas (outlined by dash line) clustered for each pathway. ( C , D ) Gene ontology (GO) analyses were performed to categorize ( C ) HIF-1α- and ( D ) HIF-2α-related genes according to their functions, in all types of ECs, including (dataset, Normal Endothelial Cells HUAEC/HUVEC - Luttun - 38 - MAS5.0 - u133p2).
Figure Legend Snippet: The GEP analyses map the potential correlations among the HIFs, TNFα, VEGFs, and NF-κB signaling pathways in ECs. ( A , B ) By analyzing the datasets involving gene expression profiling (GEP) in ECs using the R2: Genomics Analysis and Visualization Platform, the heatmaps were generated based on P values for analyses of correlations (red and blue indicating positive and negative correlations, respectively) between each pair of genes as indicated on longitudinal and transverse axes, in ( A ; dataset, Exp HUVEC vs ESC - James - 12 - MAS5.0 - u133p2) human stem cells (hESC) vs human umbilical vein endothelial cells (HUVEC) and ( B ; dataset, Normal Endothelial Cells HUAEC/HUVEC - Luttun - 38 - MAS5.0 - u133p2) HUVEC vs human umbilical artery endothelial cells (HUAEC). Numbers in the heatmaps indicate the areas (outlined by dash line) clustered for each pathway. ( C , D ) Gene ontology (GO) analyses were performed to categorize ( C ) HIF-1α- and ( D ) HIF-2α-related genes according to their functions, in all types of ECs, including (dataset, Normal Endothelial Cells HUAEC/HUVEC - Luttun - 38 - MAS5.0 - u133p2).

Techniques Used: Expressing, Generated

LDL-mediated impairment of hypoxia-induced angiogenesis involves a broad spectrum of HIF-dependent signaling pathways. ( A ) HUVECs were exposed to indicated concentrations of LDL (25 - 100 μg/ml) for 48 hrs, and then cultured under hypoxic (1% O 2 ) condition for additional 48 hrs. ( B ) HUVECs with shRNA knockdown of HIF-1α or HIF-2α were exposed to 1% O 2 (21% O 2 as normoxia control) for 24 hrs. After treatment, Western blot analysis was performed to monitor phosphorylation of AKT (S473), ERK1/2 (T202/Y204), CXCR4, VEGFR1, and VEGFR2. Blots were reprobed for β-actin as loading control. ( C ) A schematic diagram for the mechanism by which hypoxia induces angiogenesis via an autocrine loop of TNFα and resulting activation of the self-regulatory TNFα/NF-κB/HIF/VEGF signaling network in ECs, as well as the potential mechanism of action for anti-angiogenic property of LDL by which a) LDL might impair the autocrine loop of TNFα via down-regulation of its receptor TNF-R1, rather than TNFα itself; and b) LDL disrupts the TNFα-NF-κB-HIF-VEGF signaling cascade via down-regulation of multiple key components of both canonical and non-canonical NF-κB pathways.
Figure Legend Snippet: LDL-mediated impairment of hypoxia-induced angiogenesis involves a broad spectrum of HIF-dependent signaling pathways. ( A ) HUVECs were exposed to indicated concentrations of LDL (25 - 100 μg/ml) for 48 hrs, and then cultured under hypoxic (1% O 2 ) condition for additional 48 hrs. ( B ) HUVECs with shRNA knockdown of HIF-1α or HIF-2α were exposed to 1% O 2 (21% O 2 as normoxia control) for 24 hrs. After treatment, Western blot analysis was performed to monitor phosphorylation of AKT (S473), ERK1/2 (T202/Y204), CXCR4, VEGFR1, and VEGFR2. Blots were reprobed for β-actin as loading control. ( C ) A schematic diagram for the mechanism by which hypoxia induces angiogenesis via an autocrine loop of TNFα and resulting activation of the self-regulatory TNFα/NF-κB/HIF/VEGF signaling network in ECs, as well as the potential mechanism of action for anti-angiogenic property of LDL by which a) LDL might impair the autocrine loop of TNFα via down-regulation of its receptor TNF-R1, rather than TNFα itself; and b) LDL disrupts the TNFα-NF-κB-HIF-VEGF signaling cascade via down-regulation of multiple key components of both canonical and non-canonical NF-κB pathways.

Techniques Used: Cell Culture, shRNA, Western Blot, Activation Assay

18) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing

19) Product Images from "Inhibition of NF-?B activity by HIV-1 Vpr is dependent on Vpr binding protein"

Article Title: Inhibition of NF-?B activity by HIV-1 Vpr is dependent on Vpr binding protein

Journal: Journal of cellular physiology

doi: 10.1002/jcp.24226

Vpr mediated regulation of IκBα and NF-κB p65 signaling is altered by HSP27 and VprBP knockdown
Figure Legend Snippet: Vpr mediated regulation of IκBα and NF-κB p65 signaling is altered by HSP27 and VprBP knockdown

Techniques Used:

20) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane inhibits aggressiveness of hepatic carcinoma cells via the PI3K/AKT signaling pathway. (A) Expression levels of PI3K and AKT in isoflurane-treated hepatic carcinoma cells. (B) Phosphorylation levels of AKT in isoflurane-treated hepatic carcinoma cells. (C) Effects of PI3KIR on isoflurane-promoted apoptosis of hepatic carcinoma cells. Control, non-treated cells. (D) Effects of PI3KIR on survival of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Expressing

21) Product Images from "Suppressive effect mediated by human adipose-derived stem cells on T cells involves the activation of JNK"

Article Title: Suppressive effect mediated by human adipose-derived stem cells on T cells involves the activation of JNK

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2018.3953

Analysis of signaling pathways. (A) Protein levels of NF-κB P65, p-P65, IKKβ and IκBα in the total cell fraction of control and co-cultured Jurkat cells and (B) densitometric analysis of the western blot data. (C) Expression of NF-κB P65 and p-P65 in the nuclear protein of Jurkat cells and (D) densitometric analysis of the western blot data. Levels of NF-κB p-P65 were decreased in the total cell and nuclear fractions of co-cultured Jurkat cells at 72 h. NF-κB, nuclear factor-κB; p-, phosphorylated; IκBα, inhibitor of NF-κBα; IKK, IκB kinase β; J, control; co, co-cultured.
Figure Legend Snippet: Analysis of signaling pathways. (A) Protein levels of NF-κB P65, p-P65, IKKβ and IκBα in the total cell fraction of control and co-cultured Jurkat cells and (B) densitometric analysis of the western blot data. (C) Expression of NF-κB P65 and p-P65 in the nuclear protein of Jurkat cells and (D) densitometric analysis of the western blot data. Levels of NF-κB p-P65 were decreased in the total cell and nuclear fractions of co-cultured Jurkat cells at 72 h. NF-κB, nuclear factor-κB; p-, phosphorylated; IκBα, inhibitor of NF-κBα; IKK, IκB kinase β; J, control; co, co-cultured.

Techniques Used: Cell Culture, Western Blot, Expressing

22) Product Images from "Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma"

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2018.8945

Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P
Figure Legend Snippet: Isoflurane regulates aggressiveness via the PI3K/protein kinase B-mediated NF-κB signaling pathway. (A) NF-κB activity in isoflurane-pretreated hepatic carcinoma cells. (B) Expression levels of p65, IKK-β and IκBα in isoflurane-pretreated hepatic carcinoma cells. (C) Effects of PI3KIR on NF-κB activity in hepatic carcinoma cells. (D) Effects of PI3KIR on migration and invasion of hepatic carcinoma cells. Control, non-treated cells. Data are presented as the mean ± standard error of the mean of three independent experiments. **P

Techniques Used: Activity Assay, Expressing, Migration

Related Articles

Western Blot:

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma
Article Snippet: .. For western blotting, primary mouse anti-human antibodies against p65 (ab16502; 1:2,000), PI3K and (ab40776; 1:2,000), AKT (ab8805; 1:1,000), tumor necrosis factor (TNF)-α (ab6671; 1:2,000), IL-2 (cat. no. ab92381; 1:2,000), IκB kinase (IKK)-β (ab7547; 1:2,000), NF-κB inhibitor α (IκBα; ab133478; 1:2,000), pAKT (ab38449; 1:12,000) and β-actin (ab8827; 1:2,000; all from Abcam, Cambridge, UK) were added to the membranes after blocking with 5% skimmed milk for 2 h at 37°C. .. Following washing three times with PBS, membranes were incubated with secondary rabbit anti-mouse antibodies (PV-6001; 1:2,000; OriGene Technologies, Inc., Beijing, China) for 2 h at 37°C, in order to detect target proteins.

Incubation:

Article Title: Ozone therapy could attenuate tubulointerstitial injury in adenine-induced CKD rats by mediating Nrf2 and NF-κB
Article Snippet: .. The membranes were incubated with primary antibodies against Nrf2 (ab31163; Abcam, Cambridge, MA, USA), p-IκBα (sc-101713; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-NF-κB P65 (sc-33020; Santa Cruz Biotechnology, Santa Cruz, CA, USA), ICAM-1 (sc-1511; Santa Cruz Biotechnology, Santa Cruz, CA, USA), TNF-α (ab6671; Abcam, Cambridge, MA, USA), IL-1β (sc-7884; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IL-6 (sc-1266; Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4 °C. .. After washing three times with TBST, the membranes were incubated with the secondary antibody conjugated with horseradish peroxidase (ZSGB-BIO, Beijing, China) for 1 hr at room temperature.

Article Title: The Nox1/Nox4 inhibitor attenuates acute lung injury induced by ischemia-reperfusion in mice
Article Snippet: .. After embedded, sliced, deparaffinized and washed, the 4–5μm-thick sections were incubated with the rabbit polyclonal anti-NF-κBp65 antibody (catalog no ab16502, 1:500, Abcam, Cambridge,UK) and TNF-α (catalog no ab6671, 1:100, Abcam, Cambridge,UK) respectively. ..

Article Title: Ecto-5′-nucleotidase (CD73) attenuates inflammation after spinal cord injury by promoting macrophages/microglia M2 polarization in mice
Article Snippet: .. Sections were then incubated with TNF-α (1:100, Abcam, ab6671), IL-1β (1:100, Abcam, ab9722), and CD73 (1:100, Abcam, ab175396) antibodies for 1 h, followed by incubation with HRP-conjugated anti-rabbit secondary antibodies for 30 min. DAB was then added to the sections and incubated for 10 min to allow visualization of brain segments containing bound antibodies. .. All incubation processes were carried out at room temperature.

Blocking Assay:

Article Title: Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane-inhibited aggressiveness of hepatic carcinoma
Article Snippet: .. For western blotting, primary mouse anti-human antibodies against p65 (ab16502; 1:2,000), PI3K and (ab40776; 1:2,000), AKT (ab8805; 1:1,000), tumor necrosis factor (TNF)-α (ab6671; 1:2,000), IL-2 (cat. no. ab92381; 1:2,000), IκB kinase (IKK)-β (ab7547; 1:2,000), NF-κB inhibitor α (IκBα; ab133478; 1:2,000), pAKT (ab38449; 1:12,000) and β-actin (ab8827; 1:2,000; all from Abcam, Cambridge, UK) were added to the membranes after blocking with 5% skimmed milk for 2 h at 37°C. .. Following washing three times with PBS, membranes were incubated with secondary rabbit anti-mouse antibodies (PV-6001; 1:2,000; OriGene Technologies, Inc., Beijing, China) for 2 h at 37°C, in order to detect target proteins.

Article Title: The immune-microenvironment confers resistance to MAP kinase pathway inhibitors through macrophage-derived TNFα
Article Snippet: .. The TNFα blocking antibody (Ab6671, Abcam) was used at a concentration of 5ng/ml. .. Small interfering RNAs (siRNAs) were transfected using INTERFERin siRNA-transfection reagent (Polyplus, Illkirch, France) according to the manufacturer’s instructions.

Concentration Assay:

Article Title: The immune-microenvironment confers resistance to MAP kinase pathway inhibitors through macrophage-derived TNFα
Article Snippet: .. The TNFα blocking antibody (Ab6671, Abcam) was used at a concentration of 5ng/ml. .. Small interfering RNAs (siRNAs) were transfected using INTERFERin siRNA-transfection reagent (Polyplus, Illkirch, France) according to the manufacturer’s instructions.

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    Abcam nf κb iκb kinase kinase ikk β
    Analysis of signaling pathways. (A) Protein levels of NF-κB P65, p-P65, IKKβ and IκBα in the total cell fraction of control and co-cultured Jurkat cells and (B) densitometric analysis of the western blot data. (C) Expression of NF-κB P65 and p-P65 in the nuclear protein of Jurkat cells and (D) densitometric analysis of the western blot data. Levels of NF-κB p-P65 were decreased in the total cell and nuclear fractions of co-cultured Jurkat cells at 72 h. NF-κB, nuclear factor-κB; p-, phosphorylated; IκBα, inhibitor of NF-κBα; <t>IKK,</t> <t>IκB</t> kinase β; J, control; co, co-cultured.
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    Analysis of signaling pathways. (A) Protein levels of NF-κB P65, p-P65, IKKβ and IκBα in the total cell fraction of control and co-cultured Jurkat cells and (B) densitometric analysis of the western blot data. (C) Expression of NF-κB P65 and p-P65 in the nuclear protein of Jurkat cells and (D) densitometric analysis of the western blot data. Levels of NF-κB p-P65 were decreased in the total cell and nuclear fractions of co-cultured Jurkat cells at 72 h. NF-κB, nuclear factor-κB; p-, phosphorylated; IκBα, inhibitor of NF-κBα; IKK, IκB kinase β; J, control; co, co-cultured.

    Journal: International Journal of Molecular Medicine

    Article Title: Suppressive effect mediated by human adipose-derived stem cells on T cells involves the activation of JNK

    doi: 10.3892/ijmm.2018.3953

    Figure Lengend Snippet: Analysis of signaling pathways. (A) Protein levels of NF-κB P65, p-P65, IKKβ and IκBα in the total cell fraction of control and co-cultured Jurkat cells and (B) densitometric analysis of the western blot data. (C) Expression of NF-κB P65 and p-P65 in the nuclear protein of Jurkat cells and (D) densitometric analysis of the western blot data. Levels of NF-κB p-P65 were decreased in the total cell and nuclear fractions of co-cultured Jurkat cells at 72 h. NF-κB, nuclear factor-κB; p-, phosphorylated; IκBα, inhibitor of NF-κBα; IKK, IκB kinase β; J, control; co, co-cultured.

    Article Snippet: The membranes were probed with primary antibodies against phosphorylated (p)-P65 (1:2,000, cat. no. AF2006, Affinity Biosciences, Cambridge, UK), P65 (1:2,000, cat. no. 10745-1-AP, ProteinTech Group, Inc., Wuhan, China), inhibitor of NF-κB (IκB) kinase (IKK)β (1:1,000, cat. no. Ab124957, Abcam, Cambridge, MA, USA), IκBα (1:1,000, cat. no. 4812, Cell Signaling Technology, Inc., Danvers, MA, USA), B-cell lymphoma 2 (Bcl-2; 1:2,000, cat. no. 12789-1-AP, ProteinTech Group, Inc.), Bcl-2-associated X protein (Bax; 1:5,000, cat. no. 50599-2-IG, Protein Tech Group, Inc), JNK1/2 (1:1,000, cat. no. 9252, Cell Signaling Technology, Inc.), p-JNK1/2 (1:1,000, cat. no. 4668, Cell Signaling Technology, Inc.), extracellular signal-regulated kinase (ERK)1/2 (1:1,000, cat. no. 9102, Cell Signaling Technology, Inc.), p-ERK1/2 (1:2,000, cat. no. 4370, Cell Signaling Technology, Inc.), P38 (1:1,000, cat. no. 9212, Cell Signaling, Technology, Inc.), p-P38 (1:1,000, cat. no. 9211, Cell Signaling Technology, Inc.), mothers against decapentaplegic (Smad)2/3 (1:1,000, cat. no. AF6367, Affinity Biosciences), p-Smad2/3 (1:200, cat. no. MAB8935, R & D Systems, Inc.) and the loading control β-actin (1:200, cat. no. BM0627, Boster Biological Technology, Wuhan, China).

    Techniques: Cell Culture, Western Blot, Expressing