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igfbp-6 protein  (PeproTech)


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    Structured Review

    PeproTech igfbp-6 protein
    <t>IGFBP-6</t> shows higher expression in F508del-CFTR CFBE cells and is further increased after LPS treatment. (A) Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test. (B) Immunoblots of basal expression of IGFBP-6 in Wt and F508del-CFTR CFBE cells. (C) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin ( n = 3). Statistical significance tested using paired two-tailed t -test. (D,E) Cells were treated with 1 μg/ml LPS for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA s normalized first to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test (F) Immunoblots of IGFBP-6 following treatment with 1 μg/ml LPS for 24 h. (G) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin normalized to Calnexin control ( n = 3–4). * p < 0.05; ** p < 0.01; **** p < 0.0001.
    Igfbp 6 Protein, supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/igfbp-6+protein/igfbp+6+protein/pmc09322660-56-27-13
    Average 90 stars, based on 1 article reviews
    igfbp-6 protein - by Bioz Stars, 2026-11
    90/100 stars

    Images

    1) Product Images from "Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells"

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells

    Journal: Frontiers in Molecular Biosciences

    doi: 10.3389/fmolb.2022.905468

    IGFBP-6 shows higher expression in F508del-CFTR CFBE cells and is further increased after LPS treatment. (A) Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test. (B) Immunoblots of basal expression of IGFBP-6 in Wt and F508del-CFTR CFBE cells. (C) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin ( n = 3). Statistical significance tested using paired two-tailed t -test. (D,E) Cells were treated with 1 μg/ml LPS for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA s normalized first to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test (F) Immunoblots of IGFBP-6 following treatment with 1 μg/ml LPS for 24 h. (G) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin normalized to Calnexin control ( n = 3–4). * p < 0.05; ** p < 0.01; **** p < 0.0001.
    Figure Legend Snippet: IGFBP-6 shows higher expression in F508del-CFTR CFBE cells and is further increased after LPS treatment. (A) Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test. (B) Immunoblots of basal expression of IGFBP-6 in Wt and F508del-CFTR CFBE cells. (C) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin ( n = 3). Statistical significance tested using paired two-tailed t -test. (D,E) Cells were treated with 1 μg/ml LPS for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA s normalized first to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test (F) Immunoblots of IGFBP-6 following treatment with 1 μg/ml LPS for 24 h. (G) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin normalized to Calnexin control ( n = 3–4). * p < 0.05; ** p < 0.01; **** p < 0.0001.

    Techniques Used: Expressing, Quantitative RT-PCR, Two Tailed Test, Western Blot

    IGFBP-6 is reduced by Dimethyl fumarate. (A) Cells were treated with 0.1% DMSO, 1 μg/ml LPS ± 50 µM DMF or (B) PBS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) IGFBP-6 secretion after stimulation with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 24 h. Data represent the mean ± SEM ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control (Wt or F508del) for each cell line.
    Figure Legend Snippet: IGFBP-6 is reduced by Dimethyl fumarate. (A) Cells were treated with 0.1% DMSO, 1 μg/ml LPS ± 50 µM DMF or (B) PBS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) IGFBP-6 secretion after stimulation with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 24 h. Data represent the mean ± SEM ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control (Wt or F508del) for each cell line.

    Techniques Used: Quantitative RT-PCR

    Anti-IGFBP-6 antibodies increased pro-inflammatory cytokines expression. (A) F508del-CFTR CFBE cells were treated with PBS or 0.2–200 ng/ml IGFBP-6 for 4 h. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test. (B) Cells were treated with 1 μg/ml LPS ± an antibody directed against IGFBP-6 (1 μg/ml) for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNF-α mRNAs normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test.
    Figure Legend Snippet: Anti-IGFBP-6 antibodies increased pro-inflammatory cytokines expression. (A) F508del-CFTR CFBE cells were treated with PBS or 0.2–200 ng/ml IGFBP-6 for 4 h. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test. (B) Cells were treated with 1 μg/ml LPS ± an antibody directed against IGFBP-6 (1 μg/ml) for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNF-α mRNAs normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test.

    Techniques Used: Expressing, Quantitative RT-PCR, Two Tailed Test

    IGFBP-6 shows higher expression in F508del / F508del nasal epithelial cells from three CF donors and reduces pro-inflammatory cytokines expression. (A) Total RNA was extracted from 2 patients homozygous for F508del mutation and 2 non-CF donors. qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 4). Statistical significance tested using paired two-tailed t -test (B) Cells were treated with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) Cells were treated with 200 ng/ml IGFBP-6 for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNFα mRNAs normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control for (Wt or F508del) each cell line.
    Figure Legend Snippet: IGFBP-6 shows higher expression in F508del / F508del nasal epithelial cells from three CF donors and reduces pro-inflammatory cytokines expression. (A) Total RNA was extracted from 2 patients homozygous for F508del mutation and 2 non-CF donors. qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 4). Statistical significance tested using paired two-tailed t -test (B) Cells were treated with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) Cells were treated with 200 ng/ml IGFBP-6 for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNFα mRNAs normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control for (Wt or F508del) each cell line.

    Techniques Used: Expressing, Mutagenesis, Quantitative RT-PCR, Two Tailed Test

    Related Articles

    Expressing:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Quantitative RT-PCR:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Two Tailed Test:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Western Blot:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Mutagenesis:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Purification:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).

    Positive Control:

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells
    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 μM VX-661 + 3 μM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Article Title: Insulin-Like Growth Factor Binding Protein 6 Is Secreted in Extracellular Vesicles upon Hyperthermia and Oxidative Stress in Dendritic Cells But Not in Monocytes.
    Article Snippet: Purified IGFBP-6 (Peprotech, London, UK) was used a positive control (0.2 μg).



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    Image Search Results


    Effects of AAT on cytokine and antitumor factor production in CAC. Immunohistochemical staining for PCNA, β-catenin, MPO, IL-6, IGFBP-3 IGFBP-3R identified in colons from AOM/DSS-treated mice versus AOM/DSS and AAT-treated mice using an end-stage disease protocol as in . Original magnification, ×20.

    Journal: International Journal of Molecular Sciences

    Article Title: Alpha-1 Antitrypsin Inhibits Tumorigenesis and Progression of Colitis-Associated Colon Cancer through Suppression of Inflammatory Neutrophil-Activated Serine Proteases and IGFBP-3 Proteolysis

    doi: 10.3390/ijms232213737

    Figure Lengend Snippet: Effects of AAT on cytokine and antitumor factor production in CAC. Immunohistochemical staining for PCNA, β-catenin, MPO, IL-6, IGFBP-3 IGFBP-3R identified in colons from AOM/DSS-treated mice versus AOM/DSS and AAT-treated mice using an end-stage disease protocol as in . Original magnification, ×20.

    Article Snippet: Recombinant human IGFBP-3 protein was purchased from Abcam (Cambrideg, United Kingdom).

    Techniques: Immunohistochemical staining, Staining

    AAT suppresses increased serum neutrophil serine proteases PR3 and NE and subsequent IGFBP-3 proteolysis in AOM/DSS mice. ( A ) Representive WIB of sera from 4 AOM/DSS treated mice, 3 AOM/DSS+AAT treated mice and 2 control mice. AOM/DSS treated mice show increased PR3, NE and IGFBP-3 proteolysis in circulation whereas AAT administration results in significant inhibition of serum PR3, NE and increase of intact IGFBP-3 in circulation. ( B ) AAT administrated AOM/DSS mice show decreased level of oxidized-AAT but increased total AAT in circulation. ***, p < 0.001. ( C ) NSPs proteolyze IGFBP-3. Recombinant non-glycosylated human IGFBP-3 protein (30 nM) was proteolyzed by treatment with 100, 250 and 500 nM of PR3, 100 and 250 nM NE or CG for 20 min. at 37 °C. ( D – F ): AAT and AAT mimic inhibit NSP-induced IGFBP-3 proteolysis. Based upon the sequence identity among NSPs at a putative binding site of AAT and elafin, a 7 amino acid-long peptide (LIRCAML) was generated ( D ). IGFBP-3 proteolysis by PR-3 ( left figure) and NE, CG ( right figure) was inhibited by AAT (20 μM) and different concentrations of an AAT mimic (0.05, 0.1, 0.5 and 1 μM) ( E ). Few IGFBP-3 proteolytic fragments were detected in 20 μM AAT treated or 1 μM AAT mimic treated samples. Anti-protease activity of L- and D-form of N- and C-terminal modified AAT mimics ( left figure) ( F ). Nonfunctional mutant lost anti-protease activity ( right figure).

    Journal: International Journal of Molecular Sciences

    Article Title: Alpha-1 Antitrypsin Inhibits Tumorigenesis and Progression of Colitis-Associated Colon Cancer through Suppression of Inflammatory Neutrophil-Activated Serine Proteases and IGFBP-3 Proteolysis

    doi: 10.3390/ijms232213737

    Figure Lengend Snippet: AAT suppresses increased serum neutrophil serine proteases PR3 and NE and subsequent IGFBP-3 proteolysis in AOM/DSS mice. ( A ) Representive WIB of sera from 4 AOM/DSS treated mice, 3 AOM/DSS+AAT treated mice and 2 control mice. AOM/DSS treated mice show increased PR3, NE and IGFBP-3 proteolysis in circulation whereas AAT administration results in significant inhibition of serum PR3, NE and increase of intact IGFBP-3 in circulation. ( B ) AAT administrated AOM/DSS mice show decreased level of oxidized-AAT but increased total AAT in circulation. ***, p < 0.001. ( C ) NSPs proteolyze IGFBP-3. Recombinant non-glycosylated human IGFBP-3 protein (30 nM) was proteolyzed by treatment with 100, 250 and 500 nM of PR3, 100 and 250 nM NE or CG for 20 min. at 37 °C. ( D – F ): AAT and AAT mimic inhibit NSP-induced IGFBP-3 proteolysis. Based upon the sequence identity among NSPs at a putative binding site of AAT and elafin, a 7 amino acid-long peptide (LIRCAML) was generated ( D ). IGFBP-3 proteolysis by PR-3 ( left figure) and NE, CG ( right figure) was inhibited by AAT (20 μM) and different concentrations of an AAT mimic (0.05, 0.1, 0.5 and 1 μM) ( E ). Few IGFBP-3 proteolytic fragments were detected in 20 μM AAT treated or 1 μM AAT mimic treated samples. Anti-protease activity of L- and D-form of N- and C-terminal modified AAT mimics ( left figure) ( F ). Nonfunctional mutant lost anti-protease activity ( right figure).

    Article Snippet: Recombinant human IGFBP-3 protein was purchased from Abcam (Cambrideg, United Kingdom).

    Techniques: Inhibition, Recombinant, Sequencing, Binding Assay, Generated, Activity Assay, Modification, Mutagenesis

    Anti-proliferative and anti-inflammatory effects of the IGFBP-3/IGFBP-3R axis in colon cancer cells. ( A ) Basal expression of IGFBP-3R at mRNA ( top ) and protein ( bottom ) levels analyzed by Quantitative RT-PCR and WIB, respectively. The mRNA data are presented as fold of change compared to Caco-2. The values of Caco-2 were considered as 1. The cell lysates were subjected to WIB for IGFBP-3R expression. The data represent mean ± SE, n = 3 in duplicate. Heterogeneous expression of IGFBP-3R was observed. ( B ) Data demonstrating IGFBP-3-induced apoptosis in HT-29 cells ( top ). Cell death assay was performed two days after the infection. Apoptotic cell death was measured using the Cell Death Detection ELISA. The value for Ad:EV at each MOI was considered as 1. The data represent mean ± SE, n = 3 in duplicate. *, p < 0.05; **, p < 0.01; ***, p < 0.001. Representative immunoblot analysis for IGFBP-3 and IGFBP-3R after infection of Ad:IGFBP-3 ( Bottom ). Ad:EV: adenoviral plasmids with empty vector; Ad:IGFBP-3: with IGFBP-3 cDNA; Ad:IGFBP-3 GGG : with IGFBP-3 GGG mutant cDNA. Inhibitory effects of IGFBP-3 on TNF-α-induced NF-κB activity ( C ) and subsequent induction of ICAM-1 expression ( D ) in HT29 colon cells. n = 3. ( E ) Protective role of IGFBP-3 on LPS- or TNF-α-induced disruption of colonic epithelial barrier function. Measurement of TEER across confluent intestinal monolayers treated with LPS (1mM) or to TNF-α (100 ng/mL) for 2 days in the presence/absence of IGFBP-3 (30 nM) using a Millicell-ERS voltohmmeter. n = 3 in duplicate.

    Journal: International Journal of Molecular Sciences

    Article Title: Alpha-1 Antitrypsin Inhibits Tumorigenesis and Progression of Colitis-Associated Colon Cancer through Suppression of Inflammatory Neutrophil-Activated Serine Proteases and IGFBP-3 Proteolysis

    doi: 10.3390/ijms232213737

    Figure Lengend Snippet: Anti-proliferative and anti-inflammatory effects of the IGFBP-3/IGFBP-3R axis in colon cancer cells. ( A ) Basal expression of IGFBP-3R at mRNA ( top ) and protein ( bottom ) levels analyzed by Quantitative RT-PCR and WIB, respectively. The mRNA data are presented as fold of change compared to Caco-2. The values of Caco-2 were considered as 1. The cell lysates were subjected to WIB for IGFBP-3R expression. The data represent mean ± SE, n = 3 in duplicate. Heterogeneous expression of IGFBP-3R was observed. ( B ) Data demonstrating IGFBP-3-induced apoptosis in HT-29 cells ( top ). Cell death assay was performed two days after the infection. Apoptotic cell death was measured using the Cell Death Detection ELISA. The value for Ad:EV at each MOI was considered as 1. The data represent mean ± SE, n = 3 in duplicate. *, p < 0.05; **, p < 0.01; ***, p < 0.001. Representative immunoblot analysis for IGFBP-3 and IGFBP-3R after infection of Ad:IGFBP-3 ( Bottom ). Ad:EV: adenoviral plasmids with empty vector; Ad:IGFBP-3: with IGFBP-3 cDNA; Ad:IGFBP-3 GGG : with IGFBP-3 GGG mutant cDNA. Inhibitory effects of IGFBP-3 on TNF-α-induced NF-κB activity ( C ) and subsequent induction of ICAM-1 expression ( D ) in HT29 colon cells. n = 3. ( E ) Protective role of IGFBP-3 on LPS- or TNF-α-induced disruption of colonic epithelial barrier function. Measurement of TEER across confluent intestinal monolayers treated with LPS (1mM) or to TNF-α (100 ng/mL) for 2 days in the presence/absence of IGFBP-3 (30 nM) using a Millicell-ERS voltohmmeter. n = 3 in duplicate.

    Article Snippet: Recombinant human IGFBP-3 protein was purchased from Abcam (Cambrideg, United Kingdom).

    Techniques: Expressing, Quantitative RT-PCR, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Plasmid Preparation, Mutagenesis, Activity Assay

    Central hypothesis: Colonic inflammation-induced NSPs proteolyze IGFBP-3 in circulation, thereby inhibiting endocrine/paracrine/autocrine-derived antitumor and anti-inflammatory actions of IGFBP-3 in CAC. IGFBP-3 activates IGFBP-3R and induces caspase-dependent apoptosis as well as inhibiting colitis-induced NF-κB-signaling, thereby resulting in suppression of CAC progression.

    Journal: International Journal of Molecular Sciences

    Article Title: Alpha-1 Antitrypsin Inhibits Tumorigenesis and Progression of Colitis-Associated Colon Cancer through Suppression of Inflammatory Neutrophil-Activated Serine Proteases and IGFBP-3 Proteolysis

    doi: 10.3390/ijms232213737

    Figure Lengend Snippet: Central hypothesis: Colonic inflammation-induced NSPs proteolyze IGFBP-3 in circulation, thereby inhibiting endocrine/paracrine/autocrine-derived antitumor and anti-inflammatory actions of IGFBP-3 in CAC. IGFBP-3 activates IGFBP-3R and induces caspase-dependent apoptosis as well as inhibiting colitis-induced NF-κB-signaling, thereby resulting in suppression of CAC progression.

    Article Snippet: Recombinant human IGFBP-3 protein was purchased from Abcam (Cambrideg, United Kingdom).

    Techniques: Derivative Assay

    IGFBP-6 shows higher expression in F508del-CFTR CFBE cells and is further increased after LPS treatment. (A) Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test. (B) Immunoblots of basal expression of IGFBP-6 in Wt and F508del-CFTR CFBE cells. (C) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin ( n = 3). Statistical significance tested using paired two-tailed t -test. (D,E) Cells were treated with 1 μg/ml LPS for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA s normalized first to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test (F) Immunoblots of IGFBP-6 following treatment with 1 μg/ml LPS for 24 h. (G) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin normalized to Calnexin control ( n = 3–4). * p < 0.05; ** p < 0.01; **** p < 0.0001.

    Journal: Frontiers in Molecular Biosciences

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells

    doi: 10.3389/fmolb.2022.905468

    Figure Lengend Snippet: IGFBP-6 shows higher expression in F508del-CFTR CFBE cells and is further increased after LPS treatment. (A) Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test. (B) Immunoblots of basal expression of IGFBP-6 in Wt and F508del-CFTR CFBE cells. (C) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin ( n = 3). Statistical significance tested using paired two-tailed t -test. (D,E) Cells were treated with 1 μg/ml LPS for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP-6 mRNA s normalized first to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test (F) Immunoblots of IGFBP-6 following treatment with 1 μg/ml LPS for 24 h. (G) Data represent the mean ± SEM of the ratio IGFBP-6/Calnexin normalized to Calnexin control ( n = 3–4). * p < 0.05; ** p < 0.01; **** p < 0.0001.

    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 µM VX-661 + 3 µM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test, Western Blot

    IGFBP-6 is reduced by Dimethyl fumarate. (A) Cells were treated with 0.1% DMSO, 1 μg/ml LPS ± 50 µM DMF or (B) PBS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) IGFBP-6 secretion after stimulation with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 24 h. Data represent the mean ± SEM ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control (Wt or F508del) for each cell line.

    Journal: Frontiers in Molecular Biosciences

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells

    doi: 10.3389/fmolb.2022.905468

    Figure Lengend Snippet: IGFBP-6 is reduced by Dimethyl fumarate. (A) Cells were treated with 0.1% DMSO, 1 μg/ml LPS ± 50 µM DMF or (B) PBS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) IGFBP-6 secretion after stimulation with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 24 h. Data represent the mean ± SEM ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control (Wt or F508del) for each cell line.

    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 µM VX-661 + 3 µM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Techniques: Quantitative RT-PCR

    Anti-IGFBP-6 antibodies increased pro-inflammatory cytokines expression. (A) F508del-CFTR CFBE cells were treated with PBS or 0.2–200 ng/ml IGFBP-6 for 4 h. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test. (B) Cells were treated with 1 μg/ml LPS ± an antibody directed against IGFBP-6 (1 μg/ml) for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNF-α mRNAs normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test.

    Journal: Frontiers in Molecular Biosciences

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells

    doi: 10.3389/fmolb.2022.905468

    Figure Lengend Snippet: Anti-IGFBP-6 antibodies increased pro-inflammatory cytokines expression. (A) F508del-CFTR CFBE cells were treated with PBS or 0.2–200 ng/ml IGFBP-6 for 4 h. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test. (B) Cells were treated with 1 μg/ml LPS ± an antibody directed against IGFBP-6 (1 μg/ml) for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNF-α mRNAs normalized to GADPH as housekeeping gene and then to control untreated cells. Data represent the mean ± SEM ( n = 3). Statistical significance tested using paired two-tailed t -test.

    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 µM VX-661 + 3 µM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test

    IGFBP-6 shows higher expression in F508del / F508del nasal epithelial cells from three CF donors and reduces pro-inflammatory cytokines expression. (A) Total RNA was extracted from 2 patients homozygous for F508del mutation and 2 non-CF donors. qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 4). Statistical significance tested using paired two-tailed t -test (B) Cells were treated with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) Cells were treated with 200 ng/ml IGFBP-6 for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNFα mRNAs normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control for (Wt or F508del) each cell line.

    Journal: Frontiers in Molecular Biosciences

    Article Title: Insulin-Like Growth Factor Binding Protein (IGFBP-6) as a Novel Regulator of Inflammatory Response in Cystic Fibrosis Airway Cells

    doi: 10.3389/fmolb.2022.905468

    Figure Lengend Snippet: IGFBP-6 shows higher expression in F508del / F508del nasal epithelial cells from three CF donors and reduces pro-inflammatory cytokines expression. (A) Total RNA was extracted from 2 patients homozygous for F508del mutation and 2 non-CF donors. qRT-PCR was performed in order to quantify IGFBP-6 mRNA normalized to GADPH as housekeeping gene. Data represent the mean ± SEM ( n = 4). Statistical significance tested using paired two-tailed t -test (B) Cells were treated with 0.1% DMSO, 1 μg/ml LPS, 30 ng/ml IL-1β + 30 ng/ml TNFα ± 50 µM DMF for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IGFBP mRNA normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). (C) Cells were treated with 200 ng/ml IGFBP-6 for 4 h. Total RNA was extracted and qRT-PCR was performed in order to quantify IL-1β, IL-6 and TNFα mRNAs normalized to control untreated cells. Data represent the mean ± SEM ( n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significance tested using two-way ANOVA with Tukey’s multiple comparisons test to the control for (Wt or F508del) each cell line.

    Article Snippet: Cells were treated for 24 h with 0.1% DMSO or 200 ng/ml IGFBP-6 (PeproTech) or 3 µM VX-661 + 3 µM VX-445 (Selleck Chemicals) +/− 200 ng/ml IGFBP-6.

    Techniques: Expressing, Mutagenesis, Quantitative RT-PCR, Two Tailed Test