Structured Review

Cell Signaling Technology Inc icam 1
<t>ICAM-1</t> and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
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1) Product Images from "Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages"

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206759

ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
Figure Legend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Techniques Used: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

2) Product Images from "Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages"

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206759

ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
Figure Legend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Techniques Used: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

3) Product Images from "High Glucose/High Lipids Impair Vascular Adiponectin Function via Inhibition of Caveolin-1/AdipoR1 Signalsome Formation"

Article Title: High Glucose/High Lipids Impair Vascular Adiponectin Function via Inhibition of Caveolin-1/AdipoR1 Signalsome Formation

Journal: Free radical biology & medicine

doi: 10.1016/j.freeradbiomed.2015.09.005

APN attenuated TNF-α induced nitrotyrosine formation (A) and ICAM-1 expression (B) in control cells cultured with normal glucose/normal lipid (Control). After 72 hours of HUVEC exposure to HG/HL, APN failed to reduce TNF-α mediated nitrotyrosine
Figure Legend Snippet: APN attenuated TNF-α induced nitrotyrosine formation (A) and ICAM-1 expression (B) in control cells cultured with normal glucose/normal lipid (Control). After 72 hours of HUVEC exposure to HG/HL, APN failed to reduce TNF-α mediated nitrotyrosine

Techniques Used: Expressing, Cell Culture

4) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

5) Product Images from "N-acetyl-seryl-aspartyl-lysyl-proline treatment protects heart against excessive myocardial injury and heart failure in mice"

Article Title: N-acetyl-seryl-aspartyl-lysyl-proline treatment protects heart against excessive myocardial injury and heart failure in mice

Journal: Canadian journal of physiology and pharmacology

doi: 10.1139/cjpp-2019-0047

(A) Representative images of immunohistochemically stained macrophages (upper panel) and neutrophils (lower panel) in the left ventricle at 1 week of myocardial infarction (MI). (B) Quantitative analysis of CD68-positive cells (marker for macrophage), showing that N -acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) significantly reduced numbers of infiltrating macrophages. (C) There were no differences in the numbers of Ly-6b.2-positive cells (a marker for neutrophils) between the MI + vehicle and MI + Ac-SDKP groups. (D) Increased intercellular adhesion molecule-1 (ICAM-1) expression after MI was significantly inhibited by Ac-SDKP. n = 6/group, * p
Figure Legend Snippet: (A) Representative images of immunohistochemically stained macrophages (upper panel) and neutrophils (lower panel) in the left ventricle at 1 week of myocardial infarction (MI). (B) Quantitative analysis of CD68-positive cells (marker for macrophage), showing that N -acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) significantly reduced numbers of infiltrating macrophages. (C) There were no differences in the numbers of Ly-6b.2-positive cells (a marker for neutrophils) between the MI + vehicle and MI + Ac-SDKP groups. (D) Increased intercellular adhesion molecule-1 (ICAM-1) expression after MI was significantly inhibited by Ac-SDKP. n = 6/group, * p

Techniques Used: Staining, Marker, Expressing

6) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

7) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

8) Product Images from "MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection"

Article Title: MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

Journal: International journal for parasitology

doi: 10.1016/j.ijpara.2010.11.011

Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary
Figure Legend Snippet: Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary

Techniques Used: Infection, Expressing

Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following
Figure Legend Snippet: Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following

Techniques Used: Expressing, Infection, In Vitro

Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding
Figure Legend Snippet: Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding

Techniques Used: Infection, Luciferase, Construct, Binding Assay

Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced
Figure Legend Snippet: Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced

Techniques Used: Cell Culture, Functional Assay

9) Product Images from "Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages"

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206759

ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
Figure Legend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Techniques Used: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

10) Product Images from "Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages"

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206759

ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
Figure Legend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Techniques Used: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

11) Product Images from "Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages"

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206759

ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
Figure Legend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Techniques Used: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

12) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

13) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

14) Product Images from "Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction"

Article Title: Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2017.01095

LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p
Figure Legend Snippet: LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p

Techniques Used: Expressing, In Vivo, Immunohistochemistry, Staining

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing, Chick Chorioallantoic Membrane Assay

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing

15) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

16) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

17) Product Images from "MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection"

Article Title: MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

Journal: International journal for parasitology

doi: 10.1016/j.ijpara.2010.11.011

Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary
Figure Legend Snippet: Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary

Techniques Used: Infection, Expressing

Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following
Figure Legend Snippet: Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following

Techniques Used: Expressing, Infection, In Vitro

Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding
Figure Legend Snippet: Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding

Techniques Used: Infection, Luciferase, Construct, Binding Assay

Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced
Figure Legend Snippet: Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced

Techniques Used: Cell Culture, Functional Assay

18) Product Images from "MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection"

Article Title: MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

Journal: International journal for parasitology

doi: 10.1016/j.ijpara.2010.11.011

Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary
Figure Legend Snippet: Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary

Techniques Used: Infection, Expressing

Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following
Figure Legend Snippet: Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following

Techniques Used: Expressing, Infection, In Vitro

Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding
Figure Legend Snippet: Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding

Techniques Used: Infection, Luciferase, Construct, Binding Assay

Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced
Figure Legend Snippet: Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced

Techniques Used: Cell Culture, Functional Assay

19) Product Images from "MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection"

Article Title: MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

Journal: International journal for parasitology

doi: 10.1016/j.ijpara.2010.11.011

Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary
Figure Legend Snippet: Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary

Techniques Used: Infection, Expressing

Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following
Figure Legend Snippet: Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following

Techniques Used: Expressing, Infection, In Vitro

Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding
Figure Legend Snippet: Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding

Techniques Used: Infection, Luciferase, Construct, Binding Assay

Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced
Figure Legend Snippet: Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced

Techniques Used: Cell Culture, Functional Assay

20) Product Images from "Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction"

Article Title: Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2017.01095

LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p
Figure Legend Snippet: LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p

Techniques Used: Expressing, In Vivo, Immunohistochemistry, Staining

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing, Chick Chorioallantoic Membrane Assay

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing

21) Product Images from "Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction"

Article Title: Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2017.01095

LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p
Figure Legend Snippet: LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p

Techniques Used: Expressing, In Vivo, Immunohistochemistry, Staining

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing, Chick Chorioallantoic Membrane Assay

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing

22) Product Images from "The MK2/HuR signaling pathway regulates TNF-α-induced ICAM-1 expression by promoting the stabilization of ICAM-1 mRNA"

Article Title: The MK2/HuR signaling pathway regulates TNF-α-induced ICAM-1 expression by promoting the stabilization of ICAM-1 mRNA

Journal: BMC Pulmonary Medicine

doi: 10.1186/s12890-016-0247-8

Schematic model of the MK2/HuR pathway in the post-transcriptional regulation of TNF-α-induced ICAM-1 expression. The present results show that HuR nucleo-cytoplasmic shuttling mediated by TNF-α stimulation results in the stabilization of ICAM-1 mRNAs and increased protein production. The possible mechanism is that MK2 induces HuR cytoplasmic translocalization, which leads to an increase in stability of HuR-mRNA complex formation, and therefore increases ICAM-1 expression. TNFR, TNF-receptor; sICAM-1, soluble ICAM-1
Figure Legend Snippet: Schematic model of the MK2/HuR pathway in the post-transcriptional regulation of TNF-α-induced ICAM-1 expression. The present results show that HuR nucleo-cytoplasmic shuttling mediated by TNF-α stimulation results in the stabilization of ICAM-1 mRNAs and increased protein production. The possible mechanism is that MK2 induces HuR cytoplasmic translocalization, which leads to an increase in stability of HuR-mRNA complex formation, and therefore increases ICAM-1 expression. TNFR, TNF-receptor; sICAM-1, soluble ICAM-1

Techniques Used: Expressing

HuR affected the levels and stability of ICAM-1 mRNA without influencing IL-8 mRNA in HPMECs. Following TNF-α stimulation, the ICAM-1 and IL-8 mRNA levels were significantly increased in control cells, while HuR knockdown markedly reduced ICAM-1 mRNA levels but not IL-8 mRNA ( a and b ). HuR silencing decreased ICAM-1 mRNA stability, and the half-life was 118 min and 52 min in control cells and HuR-silenced cells, respectively ( c ). The half-life of IL-8 mRNA was 126 min and 114 min in control cells and HuR-silenced cells, respectively ( d ). Each point represents the mean ± SEM. n =3, # P
Figure Legend Snippet: HuR affected the levels and stability of ICAM-1 mRNA without influencing IL-8 mRNA in HPMECs. Following TNF-α stimulation, the ICAM-1 and IL-8 mRNA levels were significantly increased in control cells, while HuR knockdown markedly reduced ICAM-1 mRNA levels but not IL-8 mRNA ( a and b ). HuR silencing decreased ICAM-1 mRNA stability, and the half-life was 118 min and 52 min in control cells and HuR-silenced cells, respectively ( c ). The half-life of IL-8 mRNA was 126 min and 114 min in control cells and HuR-silenced cells, respectively ( d ). Each point represents the mean ± SEM. n =3, # P

Techniques Used:

HuR silencing reduced ICAM-1 levels, while had no effect on IL-8 expression. The Western blot showed that HuR silencing greatly reduced HuR levels and down-regulated ICAM-1 levels following TNF-α stimulation ( a and b ). ELISA analysis demonstrated a significant increase of IL-8 both in HuR-silenced cells and control cells after TNF-α stimulation, nevertheless, the IL-8 levels in both groups were statistically undistinguishable ( c ). Western blot band density and ELISA data are expressed as mean ± SEM. n =3, # P
Figure Legend Snippet: HuR silencing reduced ICAM-1 levels, while had no effect on IL-8 expression. The Western blot showed that HuR silencing greatly reduced HuR levels and down-regulated ICAM-1 levels following TNF-α stimulation ( a and b ). ELISA analysis demonstrated a significant increase of IL-8 both in HuR-silenced cells and control cells after TNF-α stimulation, nevertheless, the IL-8 levels in both groups were statistically undistinguishable ( c ). Western blot band density and ELISA data are expressed as mean ± SEM. n =3, # P

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

23) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

24) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

25) Product Images from "MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression"

Article Title: MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00355

miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p
Figure Legend Snippet: miR-200a/200b mimic decreased endothelial OGT and ICAM-1 expression in type 2 db/db diabetic mice. Immunohistochemistry staining of (A) OGT and (B) ICAMI-1 in the thoracic aorta tissue. Representative images showing that the immunoreactivities of endothelial OGT and ICAM-1 in the aortic endothelial layers (blue color as arrow) were decreased in the miR-200a/200b mimic-treated db/db mice, as compared with the negative control (NC)-treated db/db mice. db/m mice: non-diabetic control mice. N = 3 per group. Scale bar = 20 μm. (C) Immunoreactivity signals in endothelial layers from four treatment groups were quantified for all immunohistochemistry images. ** p

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Negative Control

26) Product Images from "A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase"

Article Title: A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase

Journal: Journal of Lipid Research

doi: 10.1194/jlr.M056069

Direct effects of 12/15-LOX-derived metabolites on retinal vasculature. A: Fluorescein angiography (FA) of normal WT mice injected intravitreally with vehicle, as a control, or 12-HETE. One week later, FA was performed to evaluate changes of retinal vasculature. The relative fluorescence intensity of FA per mouse retina was calculated by the ImageJ software then normalized as a percentage to that of vehicle-injected control, which was arbitrarily set at 100%. B: Western blot of total retinal albumin among the studied groups followed by densitometric analysis. Ratio of the albumin band intensity relative to actin for the 12-HETE-injected group was compared with the vehicle-injected control, which was arbitrarily set at 1.0. C: Western blot analysis of retinal ICAM-1, VCAM-1, CD45, and NOX2 after intraocular injection with either 12-HETE (0.1 µM), or vehicle followed by densitometric analysis. Ratios of band intensities of ICAM-1, VCAM-1, CD45, and NOX2, respectively, relative to the actin for 12-HETE-injected group were compared with the vehicle-injected control, which was arbitrarily set at 1.0. Data shown for the comparison are the mean ± SD and representative of four to six mice studied in each group.
Figure Legend Snippet: Direct effects of 12/15-LOX-derived metabolites on retinal vasculature. A: Fluorescein angiography (FA) of normal WT mice injected intravitreally with vehicle, as a control, or 12-HETE. One week later, FA was performed to evaluate changes of retinal vasculature. The relative fluorescence intensity of FA per mouse retina was calculated by the ImageJ software then normalized as a percentage to that of vehicle-injected control, which was arbitrarily set at 100%. B: Western blot of total retinal albumin among the studied groups followed by densitometric analysis. Ratio of the albumin band intensity relative to actin for the 12-HETE-injected group was compared with the vehicle-injected control, which was arbitrarily set at 1.0. C: Western blot analysis of retinal ICAM-1, VCAM-1, CD45, and NOX2 after intraocular injection with either 12-HETE (0.1 µM), or vehicle followed by densitometric analysis. Ratios of band intensities of ICAM-1, VCAM-1, CD45, and NOX2, respectively, relative to the actin for 12-HETE-injected group were compared with the vehicle-injected control, which was arbitrarily set at 1.0. Data shown for the comparison are the mean ± SD and representative of four to six mice studied in each group.

Techniques Used: Derivative Assay, Mouse Assay, Injection, Fluorescence, Software, Western Blot

27) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

28) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

29) Product Images from "miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes"

Article Title: miR-221 suppresses ICAM-1 translation and regulates interferon-?-induced ICAM-1 expression in human cholangiocytes

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

doi: 10.1152/ajpgi.00490.2009

IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: IFN-γ induces ICAM-1 expression through downregulation of miR-221. A : IFN-γ increased the luciferase reporter translational activity in H69 cells transfected with the construct with ICAM-1 3′UTR encoding miR-221 binding site. Cells were transfected with the pMIR-REPORT luciferase construct containing the ICAM-1 3′UTR with the putative miR-221 binding site and then exposed to IFN-γ for 24 h. Luciferase activity in cells after exposure IFN-γ was then measured and normalized to β-gal. B : miR-221 precursor blocked IFN-γ-induced ICAM-1 protein expression. H69 cells were transfected with the miR-221 precursor or a control nonspecific precursor for 48 h and then exposed to IFN-γ (10 ng/ml) for 24 h followed by Western blot for ICAM-1. C : miR-221 precursor does not alter IFN-γ-induced ICAM-1 expression at the message level. H69 cells were transfected with the miR-221 precursor for 24 h and then exposed to IFN-γ (10 ng/ml) for 8 h followed by real-time PCR analysis for ICAM-1 mRNA. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P
Figure Legend Snippet: Posttranscriptional regulation is involved in IFN-γ-induced ICAM-1 protein expression in cholangiocytes. Expression of ICAM-1 at the message ( A ) and protein ( B ) levels in H69 cells following IFN-γ stimulation in the presence or absence of actinomycin D. H69 cells were exposed to culture medium with actinomycin D (10 μg/ml) for 90 min and then exposed to IFN-γ (10 ng/ml) following by real-time PCR for ICAM-1 (after incubation for 12 h) or Western blot (after incubation for 24 h). Data are representative of 3 independent experiments. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot

Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: Interferon (IFN)-γ induces intercellular adhesion molecule (ICAM)-1 expression in cultured human cholangiocyte. A and B : dose-dependent expression of ICAM-1 at the message and the protein levels in H69 cells following IFN-γ stimulation. H69 cells were exposed to the culture medium with various doses of IFN-γ (0, 0.1, 1.0, 10, and 25 ng/ml) followed by real-time PCR (after incubation for 8 h) or Western blotting analysis for ICAM-1 (after incubation for 24 h). C and D : time-dependent expression of ICAM-1 expression in H69 cells induced by IFN-γ. Cells were exposed to IFN-γ (10 ng/ml) followed by real-time PCR (incubation for up to 24 h) or Western blot (incubation for up to 48 h). E and F : IFN-γ-induced expression of ICAM-1 in human intrahepatic biliary epithelial (HIBEpiC) cells. HIBEpiC cells were exposed to culture medium with or without IFN-γ (10 ng/ml) followed by real-time PCR (incubation for 8 h) or Western blot (incubation for 24 h). Bars represent the means ± SD from 3 independent experiments. * P

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

miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P
Figure Legend Snippet: miR-221 targets ICAM-1 3′-untranslated region (UTR) and causes posttranscriptional suppression. A : human ICAM-1 mRNA shows a potential binding site in the 3′UTR for miR-221. B : targeting of ICAM-1 3′UTR by miR-221 resulted in transcriptional suppression. The luciferase reporter constructs containing the potential binding site for miR-221 in ICAM-1 3′UTR or the mutant (Mut) sequence (TGTAGC to ACATCG) were generated. H69 cells were transiently cotransfected with the reporter construct and the miR-221 precursor or anti-miR-221 for 24 h. Luciferase activities were measured and normalized to the control (Ctrl) β-galactosidase (β-gal) level. A nonspecific precursor (precursor-Ctrl) and anti-miR (anti-miR-Ctrl) were used as the controls. Bars represent the means ± SD from 3 independent experiments. * P

Techniques Used: Binding Assay, Luciferase, Construct, Mutagenesis, Sequencing, Generated

Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P
Figure Legend Snippet: Functional inhibition of miR-221 increases ICAM-1 protein expression but does not alter ICAM-1 mRNA level. A : anti-miR-221 increased ICAM-1 protein expression in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by Western blot for ICAM-1. B : anti-miR-221 did not alter ICAM-1 expression at the message level in cholangiocytes. H69 cells were treated with various doses of anti-miR-221 for 24 h followed by real-time PCR for ICAM-1. Bars represent the means ± SD from 3 independent experiments. Anti-miR-Ctrl, nonspecific anti-miR control; * P

Techniques Used: Functional Assay, Inhibition, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P
Figure Legend Snippet: Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B : functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor ( A ) or anti-miR-221 ( B ) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C : IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. * P

Techniques Used: Functional Assay, Incubation, Labeling, Fluorescence, Microscopy

30) Product Images from "MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection"

Article Title: MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

Journal: International journal for parasitology

doi: 10.1016/j.ijpara.2010.11.011

Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary
Figure Legend Snippet: Post-transcriptional regulation is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. Expression of ICAM-1 at the message (A) and protein (B) levels in H69 human biliary

Techniques Used: Infection, Expressing

Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following
Figure Legend Snippet: Human biliary epithelial cells (H69 and HIBEpiC) up-regulate intercellular adhesion molecule-1 (ICAM-1) expression following Cryptosporidium parvum infection in vitro. (A and B) Expression of ICAM-1 at the message and protein levels in H69 cells following

Techniques Used: Expressing, Infection, In Vitro

Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding
Figure Legend Snippet: Down-regulation of microRNA-221 (miR-221) is involved in the up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in cells following Cryptosporidium parvum infection. (A) The luciferase reporter constructs containing the potential binding

Techniques Used: Infection, Luciferase, Construct, Binding Assay

Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced
Figure Legend Snippet: Up-regulation of intercellular adhesion molecule-1 (ICAM-1) protein in H69 human biliary epithelial cells induced by Cryptosporidium parvum affects adherence of co-cultured Jurkat cells. (A) Functional manipulation of microRNA-221 (miR-221) influenced

Techniques Used: Cell Culture, Functional Assay

31) Product Images from "Inhibition of interleukin-6 trans-signaling prevents inflammation and endothelial barrier disruption in retinal endothelial cells."

Article Title: Inhibition of interleukin-6 trans-signaling prevents inflammation and endothelial barrier disruption in retinal endothelial cells.

Journal: Experimental eye research

doi: 10.1016/j.exer.2018.09.009

Inhibition of IL-6 trans-signaling decreases ICAM-1 protein levels in HRECs. HRECs were treated with and without sgp130Fc (10µg/mL, 60 min) and exposed to IL-6 (10ng/mL) and sIL-6R (150ng/mL) overnight. (A) ICAM-1 expression was measured after staining with anti-ICAM-1 antibody (red). Cells were counterstained by 4′,6′-diamidino-2- phenylindole (DAPI, blue). All images were visualized by confocal microscopy. IL-6 trans-signaling activation resulted in a three-fold increase in the ICAM-1 expression. This effect was prevented by pre-treating cells with sgp130Fc. Results are expressed as means ± SEM; n = 8 per group, scale bar = 25µm, * p
Figure Legend Snippet: Inhibition of IL-6 trans-signaling decreases ICAM-1 protein levels in HRECs. HRECs were treated with and without sgp130Fc (10µg/mL, 60 min) and exposed to IL-6 (10ng/mL) and sIL-6R (150ng/mL) overnight. (A) ICAM-1 expression was measured after staining with anti-ICAM-1 antibody (red). Cells were counterstained by 4′,6′-diamidino-2- phenylindole (DAPI, blue). All images were visualized by confocal microscopy. IL-6 trans-signaling activation resulted in a three-fold increase in the ICAM-1 expression. This effect was prevented by pre-treating cells with sgp130Fc. Results are expressed as means ± SEM; n = 8 per group, scale bar = 25µm, * p

Techniques Used: Inhibition, Expressing, Staining, Confocal Microscopy, Activation Assay

32) Product Images from "Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction"

Article Title: Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2017.01095

LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p
Figure Legend Snippet: LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p

Techniques Used: Expressing, In Vivo, Immunohistochemistry, Staining

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing, Chick Chorioallantoic Membrane Assay

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing

33) Product Images from "Synergistic Inhibition of Tumor Necrosis Factor-Alpha-Stimulated Pro-Inflammatory Cytokine Expression in HaCaT Cells by a Combination of Rapamycin and Mycophenolic Acid"

Article Title: Synergistic Inhibition of Tumor Necrosis Factor-Alpha-Stimulated Pro-Inflammatory Cytokine Expression in HaCaT Cells by a Combination of Rapamycin and Mycophenolic Acid

Journal: Annals of Dermatology

doi: 10.5021/ad.2015.27.1.32

Inhibitory effects of rapamycin (RP) and mycophenolic acid (MPA) on tumor necrosis factor (TNF)-α-induced intercellular adhesion molecule 1 (ICAM-1) and inducible nitric oxide synthase (iNOS) expression in HaCaT cells. (A) HaCaT cells were pre-incubated for 24 h, and then stimulated with TNF-α (20 ng/ml) in the presence of RP, MPA, or a combination of the two drugs for 3 h (for RNA) or 12 h (for protein). ICAM-1 and iNOS expression were determined after treatment with 100 nM RP, 100 nM MPA, and different combinations of the two drugs (20 nM RP and 80 nM MPA, 50 nM RP and 50 nM MPA, and 80 nM RP and 20 nM MPA) by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. (B) HaCaT cells were pretreated with an extracellular signal-related kinases (ERK) inhibitor (PD98059), p38 inhibitor (SB203580), or c-Jun N-terminal kinases (JNK) inhibitor (SP600125) for 1 h, and then stimulated with TNF-α for 12 h. The band densities are expressed as percentages of the band density of the TNF-α-only group. ** p
Figure Legend Snippet: Inhibitory effects of rapamycin (RP) and mycophenolic acid (MPA) on tumor necrosis factor (TNF)-α-induced intercellular adhesion molecule 1 (ICAM-1) and inducible nitric oxide synthase (iNOS) expression in HaCaT cells. (A) HaCaT cells were pre-incubated for 24 h, and then stimulated with TNF-α (20 ng/ml) in the presence of RP, MPA, or a combination of the two drugs for 3 h (for RNA) or 12 h (for protein). ICAM-1 and iNOS expression were determined after treatment with 100 nM RP, 100 nM MPA, and different combinations of the two drugs (20 nM RP and 80 nM MPA, 50 nM RP and 50 nM MPA, and 80 nM RP and 20 nM MPA) by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. (B) HaCaT cells were pretreated with an extracellular signal-related kinases (ERK) inhibitor (PD98059), p38 inhibitor (SB203580), or c-Jun N-terminal kinases (JNK) inhibitor (SP600125) for 1 h, and then stimulated with TNF-α for 12 h. The band densities are expressed as percentages of the band density of the TNF-α-only group. ** p

Techniques Used: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Western Blot

34) Product Images from "Nitrosonifedipine Ameliorates the Progression of Type 2 Diabetic Nephropathy by Exerting Antioxidative Effects"

Article Title: Nitrosonifedipine Ameliorates the Progression of Type 2 Diabetic Nephropathy by Exerting Antioxidative Effects

Journal: PLoS ONE

doi: 10.1371/journal.pone.0086335

NO-NIF improved endothelial dysfunction and renal tubular injury in DN. Immunoblotting (A) and immunohistochemistry (B and C) for ICAM-1 expression in the diabetic kidney of the C57BL/6 and KKAy mice with or without NO-NIF 4 weeks after the commencement of NO-NIF (30 mg/kg) administration. (A) Representative blot of ICAM-1 and β-actin. Equal amounts of protein in each sample were separated by SDS-PAGE and analysis for ICAM-1 by western blotting. (B) Representative immunohistochemical staining of ICAM-1 in glomeruli. (C) Quantitative analysis for staining of ICAM-1 in glomeruli. Values are expressed as the means ± S.E., n = 8–10. *p
Figure Legend Snippet: NO-NIF improved endothelial dysfunction and renal tubular injury in DN. Immunoblotting (A) and immunohistochemistry (B and C) for ICAM-1 expression in the diabetic kidney of the C57BL/6 and KKAy mice with or without NO-NIF 4 weeks after the commencement of NO-NIF (30 mg/kg) administration. (A) Representative blot of ICAM-1 and β-actin. Equal amounts of protein in each sample were separated by SDS-PAGE and analysis for ICAM-1 by western blotting. (B) Representative immunohistochemical staining of ICAM-1 in glomeruli. (C) Quantitative analysis for staining of ICAM-1 in glomeruli. Values are expressed as the means ± S.E., n = 8–10. *p

Techniques Used: Immunohistochemistry, Expressing, Mouse Assay, SDS Page, Western Blot, Staining

35) Product Images from "Celastrol Inhibits Lung Infiltration in Differential Syndrome Animal Models by Reducing TNF-α and ICAM-1 Levels while Preserving Differentiation in ATRA-Induced Acute Promyelocytic Leukemia Cells"

Article Title: Celastrol Inhibits Lung Infiltration in Differential Syndrome Animal Models by Reducing TNF-α and ICAM-1 Levels while Preserving Differentiation in ATRA-Induced Acute Promyelocytic Leukemia Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0105131

Effects of celastrol on ICAM-1 expressions in ATRA-induced NB4. ( a ) Histograms of ICAM-1 expressions in NB4 cells with different treatments. NB4 cells were treated with celastrol at different doses for 30 min prior to loading 1 µM of ATRA. Cell surface levels of ICAM-1 were detected by flow cytometry at the indicated times after co-culture, with the ATRA loading time as the starting point. ( b ) Western blot detection of total amount of ICAM-1 in cells. NB4 cells were pre-treated with celastrol at 200 nM for 30 min, then 1 µM of ATRA loaded. The culture process continued for 6 hours and then the whole protein was extracted and ICAM-1 detected by Western blot. β-actin was used as internal control. ( c ) Quantitative RT-PCR detection of ICAM-1 mRNA in NB4. NB4 cells were pre-treated with celastrol at 200 nM for 30 min, then 1 µM of ATRA loaded. The culture process continued for the indicated times, after which total RNA was extracted and ICAM-1 mRNA detected with quantitative RT-PCR. β-actin was used as internal control. The data are shown as mean ± SE, * represents P
Figure Legend Snippet: Effects of celastrol on ICAM-1 expressions in ATRA-induced NB4. ( a ) Histograms of ICAM-1 expressions in NB4 cells with different treatments. NB4 cells were treated with celastrol at different doses for 30 min prior to loading 1 µM of ATRA. Cell surface levels of ICAM-1 were detected by flow cytometry at the indicated times after co-culture, with the ATRA loading time as the starting point. ( b ) Western blot detection of total amount of ICAM-1 in cells. NB4 cells were pre-treated with celastrol at 200 nM for 30 min, then 1 µM of ATRA loaded. The culture process continued for 6 hours and then the whole protein was extracted and ICAM-1 detected by Western blot. β-actin was used as internal control. ( c ) Quantitative RT-PCR detection of ICAM-1 mRNA in NB4. NB4 cells were pre-treated with celastrol at 200 nM for 30 min, then 1 µM of ATRA loaded. The culture process continued for the indicated times, after which total RNA was extracted and ICAM-1 mRNA detected with quantitative RT-PCR. β-actin was used as internal control. The data are shown as mean ± SE, * represents P

Techniques Used: Flow Cytometry, Cytometry, Co-Culture Assay, Western Blot, Quantitative RT-PCR

Relationship of a group of signaling proteins and the effects of ATRA/celastrol on ICAM-1 expressions in NB4. ( a ) The protein levels and activations for a group of signaling proteins in NB4 cells treated with ATRA or celastrol plus ATRA for different time periods. NB4 cells were pre-treated or not with celastrol for 30 min, then 1 µM ATRA added for different time periods (with ATRA addition as the starting point). The total protein was extracted and the level and phosphorylation (activation) of a group of signaling proteins were detected with respective antibodies via Western blot. ( b ) Time-dependent alterations of the ratios of p-ERK/ERK. The bands of the Western blot as shown in figure (a) were scanned and the ratios of band intensity were calculated and compared. ( c ) The effects of several signaling inhibitors and celastrol on ICAM-1 expressions in NB4. NB4 cells were pre-treated or not with celastrol or with one of the signaling inhibitors for 30 min, then 1 µM ATRA added for 24 h. The cells were then harvested and surface levels of ICAM-1 detected by flow cytometry. ( d ) The effects of signaling inhibitors on MEK1/ERK1 activation. NB4 cells were treated or not with celastrol or one of the signaling inhibitors for 30 min, and the levels and activations of MEK1 and ERK1 were detected with Western blot. The data are shown as mean ± SE, * represents P
Figure Legend Snippet: Relationship of a group of signaling proteins and the effects of ATRA/celastrol on ICAM-1 expressions in NB4. ( a ) The protein levels and activations for a group of signaling proteins in NB4 cells treated with ATRA or celastrol plus ATRA for different time periods. NB4 cells were pre-treated or not with celastrol for 30 min, then 1 µM ATRA added for different time periods (with ATRA addition as the starting point). The total protein was extracted and the level and phosphorylation (activation) of a group of signaling proteins were detected with respective antibodies via Western blot. ( b ) Time-dependent alterations of the ratios of p-ERK/ERK. The bands of the Western blot as shown in figure (a) were scanned and the ratios of band intensity were calculated and compared. ( c ) The effects of several signaling inhibitors and celastrol on ICAM-1 expressions in NB4. NB4 cells were pre-treated or not with celastrol or with one of the signaling inhibitors for 30 min, then 1 µM ATRA added for 24 h. The cells were then harvested and surface levels of ICAM-1 detected by flow cytometry. ( d ) The effects of signaling inhibitors on MEK1/ERK1 activation. NB4 cells were treated or not with celastrol or one of the signaling inhibitors for 30 min, and the levels and activations of MEK1 and ERK1 were detected with Western blot. The data are shown as mean ± SE, * represents P

Techniques Used: Activation Assay, Western Blot, Flow Cytometry, Cytometry

36) Product Images from "Interleukin-8 associates with adhesion, migration, invasion and chemosensitivity of human gastric cancer cells"

Article Title: Interleukin-8 associates with adhesion, migration, invasion and chemosensitivity of human gastric cancer cells

Journal: World Journal of Gastroenterology : WJG

doi: 10.3748/wjg.v18.i9.979

Effect of Interleukin-8 on NF-κB and Akt activities and adhesion molecules ICAM-1 and VCAM-1, and CD44 expression in gastric cancer cells. The protein expression levels of phospho-NF-κB-p65, NF-κB-p65, phospho-Akt, Akt, ICAM-1,
Figure Legend Snippet: Effect of Interleukin-8 on NF-κB and Akt activities and adhesion molecules ICAM-1 and VCAM-1, and CD44 expression in gastric cancer cells. The protein expression levels of phospho-NF-κB-p65, NF-κB-p65, phospho-Akt, Akt, ICAM-1,

Techniques Used: Expressing

37) Product Images from "FER tyrosine kinase (FER) overexpression mediates resistance to quinacrine through EGF-dependent activation of NF-?B"

Article Title: FER tyrosine kinase (FER) overexpression mediates resistance to quinacrine through EGF-dependent activation of NF-?B

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

doi: 10.1073/pnas.1105369108

Overexpression of FER activates NF-κB, and NF-κB is essential for cancer cell survival. ( A ) Overexpression of FER activates NF-κB. H1299#11 or H1299FER#11 cells were treated with quinacrine (10 μM) for 24 h, and NF-κB–dependent luciferase activity was measured 24 h later. The cell lysates were normalized for protein concentration. The results of triplicate luciferase assays are shown as means ± SD. ( B ) NF-κB is essential for cancer cell survival. RKO and H1299 cells were infected with a vector encoding a control shRNA or shRNAs against the p65 subunit of NF-κB. The cells were stained with methylene blue after infection. ( C ) The expression of p65, phospho-(S536)p65, and the NF-κB target gene ICAM-1 was analyzed by the Western method with GAPDH as a loading control.
Figure Legend Snippet: Overexpression of FER activates NF-κB, and NF-κB is essential for cancer cell survival. ( A ) Overexpression of FER activates NF-κB. H1299#11 or H1299FER#11 cells were treated with quinacrine (10 μM) for 24 h, and NF-κB–dependent luciferase activity was measured 24 h later. The cell lysates were normalized for protein concentration. The results of triplicate luciferase assays are shown as means ± SD. ( B ) NF-κB is essential for cancer cell survival. RKO and H1299 cells were infected with a vector encoding a control shRNA or shRNAs against the p65 subunit of NF-κB. The cells were stained with methylene blue after infection. ( C ) The expression of p65, phospho-(S536)p65, and the NF-κB target gene ICAM-1 was analyzed by the Western method with GAPDH as a loading control.

Techniques Used: Over Expression, Luciferase, Activity Assay, Protein Concentration, Infection, Plasmid Preparation, shRNA, Staining, Expressing, Western Blot

38) Product Images from "S-Nitrosoglutathione Protects the Spinal Bladder: Novel Therapeutic Approach to Post-Spinal Cord Injury Bladder Remodeling"

Article Title: S-Nitrosoglutathione Protects the Spinal Bladder: Novel Therapeutic Approach to Post-Spinal Cord Injury Bladder Remodeling

Journal: Neurourology and urodynamics

doi: 10.1002/nau.22619

Effect of GSNO on iNOS and ICAM-1 expression. A : Representative immunofluorescence image of bladder and kidney of experimental animals showing iNOS and ICAM-1 expression pattern. Vehicle-group had increased expression of both iNOS and ICAM-1 in both the bladder and the kidney (magnification: 200×). B : Histogram showing the number of iNOS and ICAM-1 positive cells in bladder of experimental groups. Cells were counted in 10 different regions of the sections from n = 5 in each group. C : Histogram showing the number of iNOS and ICAM-1 positive cells in the kidney of experimental animals. Cells were counted in 10 different regions of the sections from n = 5 in each group. *** P
Figure Legend Snippet: Effect of GSNO on iNOS and ICAM-1 expression. A : Representative immunofluorescence image of bladder and kidney of experimental animals showing iNOS and ICAM-1 expression pattern. Vehicle-group had increased expression of both iNOS and ICAM-1 in both the bladder and the kidney (magnification: 200×). B : Histogram showing the number of iNOS and ICAM-1 positive cells in bladder of experimental groups. Cells were counted in 10 different regions of the sections from n = 5 in each group. C : Histogram showing the number of iNOS and ICAM-1 positive cells in the kidney of experimental animals. Cells were counted in 10 different regions of the sections from n = 5 in each group. *** P

Techniques Used: Expressing, Immunofluorescence

39) Product Images from "Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction"

Article Title: Identification of Basic Fibroblast Growth Factor as the Dominant Protector of Laminar Shear Medium from the Modified Shear Device in Tumor Necrosis Factor-α Induced Endothelial Dysfunction

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2017.01095

LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p
Figure Legend Snippet: LSM and rbFGF suppresses TNF-α-induced ICAM-1 and PAI-1 protein and gene expression in vivo . (A) Immunohistochemical staining of ICAM-1 and PAI-1 proteins in thoracic aorta tissue. Representative images showing immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layers (brown color as arrow) were decreased in the TNF-α+LSM and TNF-α+rbFGF groups compared to in the TNF-α group. The immunoreactivities of ICAM-1 and PAI-1 in the aortic endothelial layer were not decreased in the TNF-α+LSM+Ab group compared to in the TNF-α group ( n = 3). (B) Immunoreactivity signals in endothelial layers from 4 groups was quantified for all immunohistochemical images. ** p

Techniques Used: Expressing, In Vivo, Immunohistochemistry, Staining

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. Expression of (A) inflammation-related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction–related HO-1, NQO-1 , and Keap-1 genes and (C) thrombosis-related TF, TM , and PAI-1 genes were all significantly increased in the TNF-α group compared to in the control group. Gene expression levels of ICAM-1, V-CAM-1, MCP-1, Keap-1, TF , and PAI-1 were attenuated significantly by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. However, gene expression of HO-1, NQO-1 , and TM were significantly enhanced by LSM treatment compared to those in the TNF-α group, but similar trends were not observed in the TNF-α+SM group. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing, Chick Chorioallantoic Membrane Assay

Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p
Figure Legend Snippet: Determination of inflammation, ROS induction, and thrombosis-related gene and protein expression. (A) Inflammation related ICAM-1, VCAM-1 , and MCP-1 genes, (B) ROS induction related Keap-1 genes, and (C) thrombosis-related TF and PAI-1 genes were attenuated significantly in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. Significant attenuation of ICAM-1, VCAM-1, MCP-1, TF , and Keap-1 were not observed in the TNF-α+LSM+Ab groups. Additionally, ROS induction-related antioxidant HO-1, NQO-1 and anti-thrombotic TM gene levels were enhanced in the TNF-α+LSM, TNF-α+LSM+IgG, and TNF-α+rbFGF groups compared to in the TNF-α groups. (A–C) Data are expressed as the mean ± S.E.M. ( n = 3). * p

Techniques Used: Expressing

40) Product Images from "The Role of TLR2 and 4-Mediated Inflammatory Pathways in Endothelial Cells Exposed to High Glucose"

Article Title: The Role of TLR2 and 4-Mediated Inflammatory Pathways in Endothelial Cells Exposed to High Glucose

Journal: PLoS ONE

doi: 10.1371/journal.pone.0108844

Effect of anti-TLR2-IgA or TAK-242 on inflammatory cytokines and cell adhesion molecules. (A) With exposure to TLR2 neutralizing antibody in control media, there was no reduction in MCP-1 and IL-8 expression however with exposure to TAK-242 in control media, there was a suppression in MCP-1 and IL-8 expression (B) With exposure to TLR2 neutralizing antibody in control media, there was no reduction in ICAM-1 expression but TAK-242 suppressed ICAM-1 expression. Normalized results are expressed as mean ± SEM, n = 3. **P
Figure Legend Snippet: Effect of anti-TLR2-IgA or TAK-242 on inflammatory cytokines and cell adhesion molecules. (A) With exposure to TLR2 neutralizing antibody in control media, there was no reduction in MCP-1 and IL-8 expression however with exposure to TAK-242 in control media, there was a suppression in MCP-1 and IL-8 expression (B) With exposure to TLR2 neutralizing antibody in control media, there was no reduction in ICAM-1 expression but TAK-242 suppressed ICAM-1 expression. Normalized results are expressed as mean ± SEM, n = 3. **P

Techniques Used: Expressing

The effect of recombinant HMGB1 on stimulating inflammatory cytokines and cell adhesion molecules. (A) Stimulating HMEC-1 cells with recombinant HMGB1 in control media for 2 hours induced the secretion of MCP-1 and IL-8 into the media (B) Exposure to recombinant HMGB1 also induced a moderate increase in ICAM-1. Normalized results are expressed as mean ± SEM, n = 3. *P
Figure Legend Snippet: The effect of recombinant HMGB1 on stimulating inflammatory cytokines and cell adhesion molecules. (A) Stimulating HMEC-1 cells with recombinant HMGB1 in control media for 2 hours induced the secretion of MCP-1 and IL-8 into the media (B) Exposure to recombinant HMGB1 also induced a moderate increase in ICAM-1. Normalized results are expressed as mean ± SEM, n = 3. *P

Techniques Used: Recombinant

Expression of cytokines and cell adhesion molecules with exposure to defined conditions for 72 hours. (A) Reduction in MCP-1 transcription was detected in the fluctuating glucose limb whereas an increase in IL-8 transcription was detected in the 30 mM glucose and fluctuating glucose limbs with maximal increase in cells exposed to fluctuating glucose concentrations for 72 hours. (B) ICAM-1 protein expression increased in both the fluctuating and 11.2 mM glucose limbs however; (C) VCAM-1 expression was not induced by any experimental condition. Normalized results are expressed as mean ± SEM, n = 4. *P
Figure Legend Snippet: Expression of cytokines and cell adhesion molecules with exposure to defined conditions for 72 hours. (A) Reduction in MCP-1 transcription was detected in the fluctuating glucose limb whereas an increase in IL-8 transcription was detected in the 30 mM glucose and fluctuating glucose limbs with maximal increase in cells exposed to fluctuating glucose concentrations for 72 hours. (B) ICAM-1 protein expression increased in both the fluctuating and 11.2 mM glucose limbs however; (C) VCAM-1 expression was not induced by any experimental condition. Normalized results are expressed as mean ± SEM, n = 4. *P

Techniques Used: Expressing

ICAM-1 expression in wildtype, TLR2 -/- and TLR4 -/- murine models. (A) In the wildtype, TLR2 -/- and TLR4 -/- murine models, ICAM-1 was expressed in the glomeruli (black arrows), peritubular capillaries (asterisks), epithelial cells (green arrows) and tubular brush border (arrowheads) (B) There was a significant upregulation of ICAM-1 in the glomeruli of wildtype diabetic mice compared to wildtype mice without diabetes. This upregulation was attenuated in TLR2 -/- and TLR4 -/- diabetic mice compared to wildtype diabetic mice. *P
Figure Legend Snippet: ICAM-1 expression in wildtype, TLR2 -/- and TLR4 -/- murine models. (A) In the wildtype, TLR2 -/- and TLR4 -/- murine models, ICAM-1 was expressed in the glomeruli (black arrows), peritubular capillaries (asterisks), epithelial cells (green arrows) and tubular brush border (arrowheads) (B) There was a significant upregulation of ICAM-1 in the glomeruli of wildtype diabetic mice compared to wildtype mice without diabetes. This upregulation was attenuated in TLR2 -/- and TLR4 -/- diabetic mice compared to wildtype diabetic mice. *P

Techniques Used: Expressing, Mouse Assay

Related Articles

Enzyme-linked Immunosorbent Assay:

Article Title: The MK2/HuR signaling pathway regulates TNF-α-induced ICAM-1 expression by promoting the stabilization of ICAM-1 mRNA
Article Snippet: .. Antibodies raised against MK2 (#3042), phospho-MK2 (Thr334, #3041), ICAM-1 (#4915) was obtained from Cell Signaling (Danvers, USA); a monoclonal antibody against HuR (ab14371) was received from Abcam (Cambridge, USA); antibodies against β-actin (P30002) and histone (P30266) were from Abmart (Shanghai, China);IL-8 ELISA Kit (BMS204/3) was received from eBioscience (San Diego, USA). .. RevertAid First Strand cDNA Synthesis Kit (#K1622) was purchased from Fermentas UAB (Vilnius, Lithuania).

Incubation:

Article Title: Vaspin inhibits cytokine-induced nuclear factor-kappa B activation and adhesion molecule expression via AMP-activated protein kinase activation in vascular endothelial cells
Article Snippet: .. Membranes were incubated in blocking buffer and then with one or more of the following primary antibodies: anti-AMPK (#2532, Cell Signaling, 1:1000), anti-phosphorylated AMPK (#2531, Thr172, Cell Signaling, 1:1000), anti-acetyl-CoA carboxylase (#3662, ACC, Cell Signaling, 1:1000), anti-phosphorylated ACC (#3661, Ser79, Cell Signaling, 1:1000), anti-phosphorylated Akt (#9271, Cell Signaling, 1:1000), anti-ICAM-1 (#4915, Cell Signaling, 1:1000), anti-VCAM-1 (#12367, Cell Signaling, 1:1000), anti-E-selectin (NBP1-45545, Novus Biologicals, CO, USA), and anti-MCP-1 (ab25124, Abcam, Cambridge, UK, 1:1000) antibodies. .. For the inhibitor of NF-κB (IκB) experiments, membranes were incubated with anti-IκBα (#4812, Cell Signaling, 1:1000), anti-phosphorylated IκBα (#4812, Ser32, Cell Signaling, 1:1000), or anti-β-actin (A5316, Sigma, St. Louis, MO, USA, 1:10,000) antibodies.

Article Title: Reduction of Endothelial Nitric Oxide Increases the Adhesiveness of Constitutive Endothelial Membrane ICAM-1 through Src-Mediated Phosphorylation
Article Snippet: .. The membranes were then incubated at room temperature in Odyssey blocking buffer (LiCor BioSciences) for 1 h, followed by incubations with primary antibodies [1:1,000 for anti-VE-Cadherin (Cat # 2500, Cell Signaling), anti ICAM-1 (1:1,000, Cat # 4915S, Cell Signaling or Cat # 554966, BD Pharmingen), and anti-β-actin (Cat # 8457, Cell Signaling), 1:500 for anti-p-ICAM-1 (phospho Y512, ab51033, Abcam)] at 4°C overnight, and secondary antibodies [1:10,000 for IRDye800-conjugated anti-mouse and anti-rabbit antibodies (Cat # 925-68071 and 925-32211, LiCor BioSciences)] at room temperature for 1 h. Images were visualized using a LiCor Odyssey Scanner and the bands were quantified using Image Studio software (LiCor BioSciences). ..

Article Title: Exosome-Transmitted miR-25 Induced by H. pylori Promotes Vascular Endothelial Cell Injury by Targeting KLF2
Article Snippet: .. The PVDF membranes were incubated with VCAM-1 (Cell Signaling Technology, CST, Boston, USA, #32653); ICAM-1 (CST, #4915); CD63 (Abcam, Shanghai, China, ab134045); KLF2 (Abcam, ab203591); or GAPDH (CST, #5174) antibodies overnight at 4°C, and then incubated with secondary antibodies (goat anti-rabbit or mouse) (ZSGB-BIO, Beijing, China) for 1 h. The PVDF membranes were then exposed in a chemiluminescence instrument (Bio-Rad ChemiDoc XRS+, USA) using the SuperSignal West Dura Extended Duration Substrate Kit (Thermo, Scientific, Beijing, China). .. Cell and Bacterial Culture The GES-1 and HEK293 cell lines were purchased from the American Type Culture Collection (ATCC).

Blocking Assay:

Article Title: Vaspin inhibits cytokine-induced nuclear factor-kappa B activation and adhesion molecule expression via AMP-activated protein kinase activation in vascular endothelial cells
Article Snippet: .. Membranes were incubated in blocking buffer and then with one or more of the following primary antibodies: anti-AMPK (#2532, Cell Signaling, 1:1000), anti-phosphorylated AMPK (#2531, Thr172, Cell Signaling, 1:1000), anti-acetyl-CoA carboxylase (#3662, ACC, Cell Signaling, 1:1000), anti-phosphorylated ACC (#3661, Ser79, Cell Signaling, 1:1000), anti-phosphorylated Akt (#9271, Cell Signaling, 1:1000), anti-ICAM-1 (#4915, Cell Signaling, 1:1000), anti-VCAM-1 (#12367, Cell Signaling, 1:1000), anti-E-selectin (NBP1-45545, Novus Biologicals, CO, USA), and anti-MCP-1 (ab25124, Abcam, Cambridge, UK, 1:1000) antibodies. .. For the inhibitor of NF-κB (IκB) experiments, membranes were incubated with anti-IκBα (#4812, Cell Signaling, 1:1000), anti-phosphorylated IκBα (#4812, Ser32, Cell Signaling, 1:1000), or anti-β-actin (A5316, Sigma, St. Louis, MO, USA, 1:10,000) antibodies.

Article Title: Reduction of Endothelial Nitric Oxide Increases the Adhesiveness of Constitutive Endothelial Membrane ICAM-1 through Src-Mediated Phosphorylation
Article Snippet: .. The membranes were then incubated at room temperature in Odyssey blocking buffer (LiCor BioSciences) for 1 h, followed by incubations with primary antibodies [1:1,000 for anti-VE-Cadherin (Cat # 2500, Cell Signaling), anti ICAM-1 (1:1,000, Cat # 4915S, Cell Signaling or Cat # 554966, BD Pharmingen), and anti-β-actin (Cat # 8457, Cell Signaling), 1:500 for anti-p-ICAM-1 (phospho Y512, ab51033, Abcam)] at 4°C overnight, and secondary antibodies [1:10,000 for IRDye800-conjugated anti-mouse and anti-rabbit antibodies (Cat # 925-68071 and 925-32211, LiCor BioSciences)] at room temperature for 1 h. Images were visualized using a LiCor Odyssey Scanner and the bands were quantified using Image Studio software (LiCor BioSciences). ..

Software:

Article Title: Reduction of Endothelial Nitric Oxide Increases the Adhesiveness of Constitutive Endothelial Membrane ICAM-1 through Src-Mediated Phosphorylation
Article Snippet: .. The membranes were then incubated at room temperature in Odyssey blocking buffer (LiCor BioSciences) for 1 h, followed by incubations with primary antibodies [1:1,000 for anti-VE-Cadherin (Cat # 2500, Cell Signaling), anti ICAM-1 (1:1,000, Cat # 4915S, Cell Signaling or Cat # 554966, BD Pharmingen), and anti-β-actin (Cat # 8457, Cell Signaling), 1:500 for anti-p-ICAM-1 (phospho Y512, ab51033, Abcam)] at 4°C overnight, and secondary antibodies [1:10,000 for IRDye800-conjugated anti-mouse and anti-rabbit antibodies (Cat # 925-68071 and 925-32211, LiCor BioSciences)] at room temperature for 1 h. Images were visualized using a LiCor Odyssey Scanner and the bands were quantified using Image Studio software (LiCor BioSciences). ..

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    Cell Signaling Technology Inc icam 1
    <t>ICAM-1</t> and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p
    Icam 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icam 1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    icam 1 - by Bioz Stars, 2020-09
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    91
    Cell Signaling Technology Inc antibody against icam 1
    p-STAT3 upregulates <t>ICAM-1</t> expression in GSC11 cells under hypoxic conditions ( A ) Western blotting revealed that ICAM-1 is expressed in multiple human glioma stem cell lines. ( B ) Western blotting revealed that the ICAM-1 expression level in GSC11 cells increased in a time-dependent manner under hypoxic conditions. ( C ) Western blotting revealed that ICAM-1 was expressed in GSC11 cells treated with or without LY, U0126, SU, or AZD1480 under hypoxic conditions. ( D ) Western blotting revealed the p-STAT3 expression level in the nuclear fraction of GSC11 cells treated with or without AZD1480 under hypoxic conditions.
    Antibody Against Icam 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against icam 1/product/Cell Signaling Technology Inc
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    antibody against icam 1 - by Bioz Stars, 2020-09
    91/100 stars
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    Image Search Results


    ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

    Journal: PLoS ONE

    Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

    doi: 10.1371/journal.pone.0206759

    Figure Lengend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

    Article Snippet: When only THP-1-derived macrophages (no exosomes) were visualized, little ICAM-1 signal was observed, suggesting that the ICAM-1 was mainly associated with the AsPC-1 exosomes and not with the THP-1 macrophages, and the CD11c signal remained at the cell surface.

    Techniques: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

    APN attenuated TNF-α induced nitrotyrosine formation (A) and ICAM-1 expression (B) in control cells cultured with normal glucose/normal lipid (Control). After 72 hours of HUVEC exposure to HG/HL, APN failed to reduce TNF-α mediated nitrotyrosine

    Journal: Free radical biology & medicine

    Article Title: High Glucose/High Lipids Impair Vascular Adiponectin Function via Inhibition of Caveolin-1/AdipoR1 Signalsome Formation

    doi: 10.1016/j.freeradbiomed.2015.09.005

    Figure Lengend Snippet: APN attenuated TNF-α induced nitrotyrosine formation (A) and ICAM-1 expression (B) in control cells cultured with normal glucose/normal lipid (Control). After 72 hours of HUVEC exposure to HG/HL, APN failed to reduce TNF-α mediated nitrotyrosine

    Article Snippet: Antibodies against APN receptor 1 (AdipoR1), APN receptor 2 (AdipoR2), endothelial NO synthase (eNOS), AMP activated protein kinase (AMPK), Akt, ICAM-1, GAPDH, eNOS phosphorylated at Ser1177, phosphorylated AMPK, and phosphorylated Akt were from Cell Signaling Technology (Beverly, MA).

    Techniques: Expressing, Cell Culture

    p-STAT3 upregulates ICAM-1 expression in GSC11 cells under hypoxic conditions ( A ) Western blotting revealed that ICAM-1 is expressed in multiple human glioma stem cell lines. ( B ) Western blotting revealed that the ICAM-1 expression level in GSC11 cells increased in a time-dependent manner under hypoxic conditions. ( C ) Western blotting revealed that ICAM-1 was expressed in GSC11 cells treated with or without LY, U0126, SU, or AZD1480 under hypoxic conditions. ( D ) Western blotting revealed the p-STAT3 expression level in the nuclear fraction of GSC11 cells treated with or without AZD1480 under hypoxic conditions.

    Journal: Oncotarget

    Article Title: Targeting intercellular adhesion molecule-1 prolongs survival in mice bearing bevacizumab-resistant glioblastoma

    doi: 10.18632/oncotarget.18859

    Figure Lengend Snippet: p-STAT3 upregulates ICAM-1 expression in GSC11 cells under hypoxic conditions ( A ) Western blotting revealed that ICAM-1 is expressed in multiple human glioma stem cell lines. ( B ) Western blotting revealed that the ICAM-1 expression level in GSC11 cells increased in a time-dependent manner under hypoxic conditions. ( C ) Western blotting revealed that ICAM-1 was expressed in GSC11 cells treated with or without LY, U0126, SU, or AZD1480 under hypoxic conditions. ( D ) Western blotting revealed the p-STAT3 expression level in the nuclear fraction of GSC11 cells treated with or without AZD1480 under hypoxic conditions.

    Article Snippet: Blots were incubated with the primary antibody against ICAM-1 (1:1000; Cell signaling Technology, Danvers, MA), GFP (1:1000, CST), p-STAT3 (1:1000, Cell signaling), Tubulin (1:3000; Sigma), p-AKT (1: 1000, Cell signaling Technology, Danvers, MA), AKT (1: 1000, Cell signaling).

    Techniques: Expressing, Western Blot

    ICAM-1 knockdown inhibits macrophage infiltration into tumor in bevacizumab-treated mice ( A ) and ( B ) Immunofluorescence staining with F4/80 (red) (A) and the bar graph of F4/80 positive cells in each high powered microscopic field (B) revealed that shRNA ICAM-1generated tumor had less F4/80 positive macrophage infiltration compared with GSC11GFP generated tumor. Representative photomicrographic images are shown (magnification ×200). ( C ) Immunofluorescence staining for Nestin (green) and F4/80 revealed that blocking ICAM-1 by treatment with ICAM-1 antibody 10 µg/ml or 20 µg/ml in GSC11 cells reduced GSC11 binding to macrophages. ( D ) The bar graph represents the ratio of Nestin positive to F4/80 positive. Data showed that blocking ICAM-1 decreased GSC capacity of binding to macrophages.

    Journal: Oncotarget

    Article Title: Targeting intercellular adhesion molecule-1 prolongs survival in mice bearing bevacizumab-resistant glioblastoma

    doi: 10.18632/oncotarget.18859

    Figure Lengend Snippet: ICAM-1 knockdown inhibits macrophage infiltration into tumor in bevacizumab-treated mice ( A ) and ( B ) Immunofluorescence staining with F4/80 (red) (A) and the bar graph of F4/80 positive cells in each high powered microscopic field (B) revealed that shRNA ICAM-1generated tumor had less F4/80 positive macrophage infiltration compared with GSC11GFP generated tumor. Representative photomicrographic images are shown (magnification ×200). ( C ) Immunofluorescence staining for Nestin (green) and F4/80 revealed that blocking ICAM-1 by treatment with ICAM-1 antibody 10 µg/ml or 20 µg/ml in GSC11 cells reduced GSC11 binding to macrophages. ( D ) The bar graph represents the ratio of Nestin positive to F4/80 positive. Data showed that blocking ICAM-1 decreased GSC capacity of binding to macrophages.

    Article Snippet: Blots were incubated with the primary antibody against ICAM-1 (1:1000; Cell signaling Technology, Danvers, MA), GFP (1:1000, CST), p-STAT3 (1:1000, Cell signaling), Tubulin (1:3000; Sigma), p-AKT (1: 1000, Cell signaling Technology, Danvers, MA), AKT (1: 1000, Cell signaling).

    Techniques: Mouse Assay, Immunofluorescence, Staining, shRNA, Generated, Blocking Assay, Binding Assay

    ICAM-1 is overexpressed in bevacizumab-resistant GSC11 xenografts ( A ) Quantitative real-time PCR, in which GAPDH mRNA expression was used as internal control, revealed that the fold-change in the RNA expression level of ICAM-1 in xenografts from bevacizumab-treated mice was significantly higher than that in vehicle-treated control mice. ( B ) Immunohistochemical analysis revealed that GSC11 xenografts from mice treated with vehicle only (Control) had lower ICAM-1 expression than those from mice treated with bevacizumab (Bev) did. Representative images are shown; cell membranes expressing ICAM-1 are brown (indicated by red arrows; magnification × 400). ( C ) Western blotting revealed that ICAM-1 was expressed in the GSC11 xenografts regardless of whether mice were treated with vehicle (Control) or bevacizumab (Bev).

    Journal: Oncotarget

    Article Title: Targeting intercellular adhesion molecule-1 prolongs survival in mice bearing bevacizumab-resistant glioblastoma

    doi: 10.18632/oncotarget.18859

    Figure Lengend Snippet: ICAM-1 is overexpressed in bevacizumab-resistant GSC11 xenografts ( A ) Quantitative real-time PCR, in which GAPDH mRNA expression was used as internal control, revealed that the fold-change in the RNA expression level of ICAM-1 in xenografts from bevacizumab-treated mice was significantly higher than that in vehicle-treated control mice. ( B ) Immunohistochemical analysis revealed that GSC11 xenografts from mice treated with vehicle only (Control) had lower ICAM-1 expression than those from mice treated with bevacizumab (Bev) did. Representative images are shown; cell membranes expressing ICAM-1 are brown (indicated by red arrows; magnification × 400). ( C ) Western blotting revealed that ICAM-1 was expressed in the GSC11 xenografts regardless of whether mice were treated with vehicle (Control) or bevacizumab (Bev).

    Article Snippet: Blots were incubated with the primary antibody against ICAM-1 (1:1000; Cell signaling Technology, Danvers, MA), GFP (1:1000, CST), p-STAT3 (1:1000, Cell signaling), Tubulin (1:3000; Sigma), p-AKT (1: 1000, Cell signaling Technology, Danvers, MA), AKT (1: 1000, Cell signaling).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, RNA Expression, Mouse Assay, Immunohistochemistry, Western Blot

    ICAM-1 knockdown prolongs survival in mice with bevacizumab-resistant glioblastoma ( A ) Kaplan-Meier survival analysis revealed that when mice bearing shRNA ICAM-1 #4 xenografts prolonger survival compared with the mice bearing GSC11 GFP after treatment with bevacizumab. H E staining analysis ( B ) and the bar graph of tumor volume ( C ) revealed that the size of the tumor which from mice bearing shRNA ICAM-1 #4 xenografts significantly decreased compared with the tumor from the mice bearing GSC11 GFP after with or without treatment of bevacizumab for 5 weeks. Representative images are shown ( D ) for the immunofluorescence staining of ICAM-1 (green) revealing that ICAM-1 expression was significantly lower in tumor from mice bearing shRNA ICAM-1 xenografts compared with tumor from mice bearing GSC11GFP after treatment with or without bevacizumab. Representative photomicrograph images are shown (magnification × 200). ( E ) The bar graph from the percentage of ICAM-1 staining in tumor area (D) revealed that ICAM-1 expression was blocked in tumor from mice bearing shRNA ICAM-1#4 compared with tumor from the mice bearing GSC11 GFP ( * P

    Journal: Oncotarget

    Article Title: Targeting intercellular adhesion molecule-1 prolongs survival in mice bearing bevacizumab-resistant glioblastoma

    doi: 10.18632/oncotarget.18859

    Figure Lengend Snippet: ICAM-1 knockdown prolongs survival in mice with bevacizumab-resistant glioblastoma ( A ) Kaplan-Meier survival analysis revealed that when mice bearing shRNA ICAM-1 #4 xenografts prolonger survival compared with the mice bearing GSC11 GFP after treatment with bevacizumab. H E staining analysis ( B ) and the bar graph of tumor volume ( C ) revealed that the size of the tumor which from mice bearing shRNA ICAM-1 #4 xenografts significantly decreased compared with the tumor from the mice bearing GSC11 GFP after with or without treatment of bevacizumab for 5 weeks. Representative images are shown ( D ) for the immunofluorescence staining of ICAM-1 (green) revealing that ICAM-1 expression was significantly lower in tumor from mice bearing shRNA ICAM-1 xenografts compared with tumor from mice bearing GSC11GFP after treatment with or without bevacizumab. Representative photomicrograph images are shown (magnification × 200). ( E ) The bar graph from the percentage of ICAM-1 staining in tumor area (D) revealed that ICAM-1 expression was blocked in tumor from mice bearing shRNA ICAM-1#4 compared with tumor from the mice bearing GSC11 GFP ( * P

    Article Snippet: Blots were incubated with the primary antibody against ICAM-1 (1:1000; Cell signaling Technology, Danvers, MA), GFP (1:1000, CST), p-STAT3 (1:1000, Cell signaling), Tubulin (1:3000; Sigma), p-AKT (1: 1000, Cell signaling Technology, Danvers, MA), AKT (1: 1000, Cell signaling).

    Techniques: Mouse Assay, shRNA, Staining, Immunofluorescence, Expressing

    ICAM-1 knockdown suppresses cell invasion in vitro and in vivo ( A ) and ( B ) GSC11GFP control and shRNA ICAM-1#4 cells treated with or without bevacizumab were subjected to a transwell migration assay. Photomicrographs of representative samples from the assay (A) and the bar graph of quantitative absorbance (590 nm) (B) revealed that shRNA ICAM-1#2 and shRNA ICAM-1 #4 cells significantly were less invaded compared with GSC GFP after cells treated with or without bevacizumab. Representative photomicrographs from three independent experiments are shown (magnification ×200) ( * P

    Journal: Oncotarget

    Article Title: Targeting intercellular adhesion molecule-1 prolongs survival in mice bearing bevacizumab-resistant glioblastoma

    doi: 10.18632/oncotarget.18859

    Figure Lengend Snippet: ICAM-1 knockdown suppresses cell invasion in vitro and in vivo ( A ) and ( B ) GSC11GFP control and shRNA ICAM-1#4 cells treated with or without bevacizumab were subjected to a transwell migration assay. Photomicrographs of representative samples from the assay (A) and the bar graph of quantitative absorbance (590 nm) (B) revealed that shRNA ICAM-1#2 and shRNA ICAM-1 #4 cells significantly were less invaded compared with GSC GFP after cells treated with or without bevacizumab. Representative photomicrographs from three independent experiments are shown (magnification ×200) ( * P

    Article Snippet: Blots were incubated with the primary antibody against ICAM-1 (1:1000; Cell signaling Technology, Danvers, MA), GFP (1:1000, CST), p-STAT3 (1:1000, Cell signaling), Tubulin (1:3000; Sigma), p-AKT (1: 1000, Cell signaling Technology, Danvers, MA), AKT (1: 1000, Cell signaling).

    Techniques: In Vitro, In Vivo, shRNA, Transwell Migration Assay