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    ProSci Incorporated irak m
    Irak M, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam anti irak3 antibody
    qRT-PCR Primers
    Anti Irak3 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam anti irak3 antibodies
    qRT-PCR Primers
    Anti Irak3 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    irak3  (Abcam)
    86
    Abcam irak3
    <t>IRAK3</t> was a downstream target of miR-591. a The predicted binding sites of miR-591 in the 3′UTR of IRAK3 mRNA were identified by using the starBase database and the relative luciferase activity in chondrocytes co-transfected with wt-IRAK3 or mut-IRAK3 and miR-591 mimic or miRNA NC was determined by dual-luciferase reporter assay. b The level of miR-591 was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. c The level of IRAK3 protein expression level was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. d The IRAK3 mRNA expression in OA cartilage and normal cartilage. e The IRAK3 protein expression in OA cartilage and normal cartilage. * P <0.05
    Irak3, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam rabbit polyclonal anti irak 3 antibody
    A) Fold changes in IRAK3 gene expression in healthy lungs after 4 hours of spontaneous breathing (non-ventilated) or mechanical ventilation with 6 ml/kg and 20 ml/kg . Bars represent the median fold-increase compared to non-ventilated animals. (*) p = 0.001 vs . non-ventilated animals. B) Representative blots from individual animals showing changes of <t>IRAK-3</t> protein levels in lungs after 4 hours of mechanical ventilation with low or high V T . Histograms represent mean densitometric values showing IRAK-3 protein levels from all animals in each group (n = 6 animals per group). Data are reported as means ± SD and were obtained from 6 independent experiments. (*) p = 0.001 vs . non-ventilated animals. V T = tidal volume.
    Rabbit Polyclonal Anti Irak 3 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    qRT-PCR Primers

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: qRT-PCR Primers

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Sequencing

    Methylation-Specific Primers

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Methylation-Specific Primers

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Methylation, Sequencing

    DNA methylation analysis of tumor tissues and normal tissues and analysis of methylation degree for seven CpG sites of IRAK3 . ( A ) The expression of the top 20 candidate genes was analyzed. IRAK3 was hypermethylated significantly in tumor tissues compared with normal tissues. ( B ) The number of IRAK3 –methylated CpG islands in each isosite. ( C – I ) Seven CpG sites for IRAK3 , which are presented in the boxplot, displayed a decreased methylation in the tumor group. The boxplot for cg 26,415,547 ( C ), cg 26,279,550 ( D ), cg 20,162,652 ( E ), cg 18,177,616 ( F ), cg 13,807,985 ( G ), cg 10,389,229 ( H ), and cg 01263292 ( I ). Abbreviation: MG, malignant glioma.

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: DNA methylation analysis of tumor tissues and normal tissues and analysis of methylation degree for seven CpG sites of IRAK3 . ( A ) The expression of the top 20 candidate genes was analyzed. IRAK3 was hypermethylated significantly in tumor tissues compared with normal tissues. ( B ) The number of IRAK3 –methylated CpG islands in each isosite. ( C – I ) Seven CpG sites for IRAK3 , which are presented in the boxplot, displayed a decreased methylation in the tumor group. The boxplot for cg 26,415,547 ( C ), cg 26,279,550 ( D ), cg 20,162,652 ( E ), cg 18,177,616 ( F ), cg 13,807,985 ( G ), cg 10,389,229 ( H ), and cg 01263292 ( I ). Abbreviation: MG, malignant glioma.

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: DNA Methylation Assay, Methylation, Expressing

    Analysis of the IRAK3 -related signaling pathway. ( A ) The top 7 signaling pathways with the highest and lowest correlation with glioma. ( B ) The top 30 signaling pathway–related genes enriched in the glioma; the MAPK signaling pathway was highly expressed. ( C ) The MAPK signaling pathway was activated in glioma. ( D ) The status of the top 16 enriched signaling pathways in glioma. Abbreviation: MG, malignant glioma.

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Analysis of the IRAK3 -related signaling pathway. ( A ) The top 7 signaling pathways with the highest and lowest correlation with glioma. ( B ) The top 30 signaling pathway–related genes enriched in the glioma; the MAPK signaling pathway was highly expressed. ( C ) The MAPK signaling pathway was activated in glioma. ( D ) The status of the top 16 enriched signaling pathways in glioma. Abbreviation: MG, malignant glioma.

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques:

    Analysis of the IRAK3 -related gene expression level. ( A ) Based on the differential genes in the disease data, the distribution of upregulated genes and downregulated genes in the GO-related pathway was enriched. ( B ) The expression level of top 40 MAPK signaling pathway–related genes. Abbreviation: MG, malignant glioma.

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Analysis of the IRAK3 -related gene expression level. ( A ) Based on the differential genes in the disease data, the distribution of upregulated genes and downregulated genes in the GO-related pathway was enriched. ( B ) The expression level of top 40 MAPK signaling pathway–related genes. Abbreviation: MG, malignant glioma.

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Expressing

    Methylation level and expression level of the IRAK3 in glioma tissues and cells. ( A ) The IRAK3 was highly methylated in glioma tissues compared with adjacent tissues. ( B ) The IRAK3 was less expressed in glioma tissues than in adjacent tissues. ( C ) The IRAK3 in the five glioma cells (SHG-44, U251, HS683, SF-539, and GOS-3) was hypermethylated compared with U251 glioma cells. ( D ) The IRAK3 was less expressed in the five glioma cells than in normal glioma cells. The difference was significant (** P < 0.01).

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Methylation level and expression level of the IRAK3 in glioma tissues and cells. ( A ) The IRAK3 was highly methylated in glioma tissues compared with adjacent tissues. ( B ) The IRAK3 was less expressed in glioma tissues than in adjacent tissues. ( C ) The IRAK3 in the five glioma cells (SHG-44, U251, HS683, SF-539, and GOS-3) was hypermethylated compared with U251 glioma cells. ( D ) The IRAK3 was less expressed in the five glioma cells than in normal glioma cells. The difference was significant (** P < 0.01).

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Methylation, Expressing

    Effect on glioma cells after the overexpression and demethylation of the IRAK3 . ( A ) The expression level of the IRAK3 was significantly higher in the glioma U251 cells of the pcDNA3.1-IRAK3 and 5-aza-dC groups than in the control group. ( B and C ) The protein expression level of the IRAK3 significantly increased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( D ) The expression level of the inflammatory factor IL-6 significantly decreased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( E ) The activity significantly decreased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( F and G ) The apoptosis rate significantly increased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( H and I ) The migration capability was significantly lower in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( J and K ) The invasiveness capability was significantly lower in the pcDNA3.1-IRAK3 and 5-aza-dC groups than in the the control group. All the mentioned differences were significant (** P < 0.01). The IRAK3 had a suppressive effect on glioma cells in vitro.

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Effect on glioma cells after the overexpression and demethylation of the IRAK3 . ( A ) The expression level of the IRAK3 was significantly higher in the glioma U251 cells of the pcDNA3.1-IRAK3 and 5-aza-dC groups than in the control group. ( B and C ) The protein expression level of the IRAK3 significantly increased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( D ) The expression level of the inflammatory factor IL-6 significantly decreased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( E ) The activity significantly decreased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( F and G ) The apoptosis rate significantly increased in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( H and I ) The migration capability was significantly lower in the pcDNA3.1-IRAK3 and 5-aza-dC groups compared with the control group. ( J and K ) The invasiveness capability was significantly lower in the pcDNA3.1-IRAK3 and 5-aza-dC groups than in the the control group. All the mentioned differences were significant (** P < 0.01). The IRAK3 had a suppressive effect on glioma cells in vitro.

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Over Expression, Expressing, Activity Assay, Migration, In Vitro

    Effect on glioma cells after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A ) The expression level of MAPK signaling pathway–related genes expression level was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( B and C ) The expression level of MAPK signaling pathway–related proteins significantly decreased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. ( D ) The level of inflammatory factor IL-6 was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( E ) The activity significantly decreased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. ( F and G ) The apoptosis rate significantly increased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. All the mentioned differences were significant (* P < 0.05, ** P < 0.01).

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Effect on glioma cells after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A ) The expression level of MAPK signaling pathway–related genes expression level was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( B and C ) The expression level of MAPK signaling pathway–related proteins significantly decreased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. ( D ) The level of inflammatory factor IL-6 was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( E ) The activity significantly decreased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. ( F and G ) The apoptosis rate significantly increased in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups compared with the control group. All the mentioned differences were significant (* P < 0.05, ** P < 0.01).

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Expressing, Activity Assay

    Effect on the migration and invasiveness capability of glioma cells after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A and B ) The migration capability was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( A and C ) The invasiveness capability was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. All the mentioned differences were significant (** P < 0.01).

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Effect on the migration and invasiveness capability of glioma cells after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A and B ) The migration capability was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. ( A and C ) The invasiveness capability was significantly lower in the PD325901, pcDNA3.1-IRAK3, and 5-aza-dC groups than in the control group. All the mentioned differences were significant (** P < 0.01).

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Migration

    Effect on mice after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A ) The tumor in the transplantation PD325901, p- IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups after 7, 14, 21, and 28 days. ( B ) The tumor volume was smaller in the PD325901, pcDNA3.1-IRAK3+PD325901, and 5-aza-dC+PD325901 groups compared with the control group. ( C ) The tumor weight followed the same trend as the volume. ( D ) The expression level of the IRAK3 was lower in the transplantation PD325901, pcDNA3.1-IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups compared with the transplantation control group. ( E and F ) The protein expression level of IRAK3 was lower in the transplantation PD325901, pcDNA3.1-IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups than in the transplantation control group. All the mentioned differences were significant (* P < 0.05, ** P < 0.01).

    Journal: Cancer Management and Research

    Article Title: Hypermethylation of the IRAK3 -Activated MAPK Signaling Pathway to Promote the Development of Glioma

    doi: 10.2147/CMAR.S252772

    Figure Lengend Snippet: Effect on mice after treatment with the MAPK signaling pathway inhibitor PD325901, pcDNA3.1-IRAK3, and 5-aza-dC. ( A ) The tumor in the transplantation PD325901, p- IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups after 7, 14, 21, and 28 days. ( B ) The tumor volume was smaller in the PD325901, pcDNA3.1-IRAK3+PD325901, and 5-aza-dC+PD325901 groups compared with the control group. ( C ) The tumor weight followed the same trend as the volume. ( D ) The expression level of the IRAK3 was lower in the transplantation PD325901, pcDNA3.1-IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups compared with the transplantation control group. ( E and F ) The protein expression level of IRAK3 was lower in the transplantation PD325901, pcDNA3.1-IRAK3 + PD325901, and 5-aza-dC + PD325901 treatment groups than in the transplantation control group. All the mentioned differences were significant (* P < 0.05, ** P < 0.01).

    Article Snippet: The membranes were probed with primary anti- IRAK3 antibody (ab8116, Abcam, MA, USA), anti- MEK1 (phospho S298) antibody [EPR3338] (ab96379), anti- ERK1 + ERK2 antibody [ERK-7D8] (ab54230), anti-c- Fos (phospho T232) antibody (ab17933), and anti- GAPDH antibody [6C5] (ab8245, Abcam) as control.

    Techniques: Transplantation Assay, Expressing

    IRAK3 was a downstream target of miR-591. a The predicted binding sites of miR-591 in the 3′UTR of IRAK3 mRNA were identified by using the starBase database and the relative luciferase activity in chondrocytes co-transfected with wt-IRAK3 or mut-IRAK3 and miR-591 mimic or miRNA NC was determined by dual-luciferase reporter assay. b The level of miR-591 was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. c The level of IRAK3 protein expression level was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. d The IRAK3 mRNA expression in OA cartilage and normal cartilage. e The IRAK3 protein expression in OA cartilage and normal cartilage. * P <0.05

    Journal: Journal of Orthopaedic Surgery and Research

    Article Title: Knockdown of Circ_SLC39A8 protects against the progression of osteoarthritis by regulating miR-591/IRAK3 axis

    doi: 10.1186/s13018-021-02323-7

    Figure Lengend Snippet: IRAK3 was a downstream target of miR-591. a The predicted binding sites of miR-591 in the 3′UTR of IRAK3 mRNA were identified by using the starBase database and the relative luciferase activity in chondrocytes co-transfected with wt-IRAK3 or mut-IRAK3 and miR-591 mimic or miRNA NC was determined by dual-luciferase reporter assay. b The level of miR-591 was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. c The level of IRAK3 protein expression level was determined in OA chondrocytes transfected with NC, miR-591, anti-NC, or anti-miR-591. d The IRAK3 mRNA expression in OA cartilage and normal cartilage. e The IRAK3 protein expression in OA cartilage and normal cartilage. * P <0.05

    Article Snippet: Next, the membranes were blocked with 5% non-fat milk at room temperature for 2 h and incubated overnight at 4 °C with primary antibodies: Bcl-2 (Abcam, 1:500), Bax (Abcam, 1:500), caspase-3 (Abcam, 1:500), cleaved caspase-3 (Abcam, 1:500), caspase-9 (Abcam, 1:500), cleaved caspase-9 (Abcam, 1:500), GAPDH (Abcam, 1:500), TNF-α (Abcam, 1:500), IL-1β (Abcam, 1:500), IL-6 (Abcam, 1:500), MMP-3 (Abcam, 1:500), COL2A1 (Abcam, 1:500), and IRAK3 (Abcam, 1:500).

    Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Reporter Assay, Expressing

    Overexpression of IRAK3 partially reversed miR-591 on chondrocyte apoptosis. a Overexpression efficiency of IRAK3 was determined by western blot assay in OA chondrocytes transfected with IRAK3 or vector. b IRAK expression in OA chondrocytes transfected with NC, miR-591, miR-591 + vector, or miR-591 + IRAK3. c Cell apoptosis was determined using flow cytometry analysis. d The protein levels of Bcl-2, Bax, caspase 3, cleaved-caspase 3, caspase 9, and cleaved-caspase 9 were analyzed by western blot assay. e Western blot assay was carried out to examine the protein expression of TNF-α, IL-1β, IL-6, MMP3, and COL2A1. * P <0.05

    Journal: Journal of Orthopaedic Surgery and Research

    Article Title: Knockdown of Circ_SLC39A8 protects against the progression of osteoarthritis by regulating miR-591/IRAK3 axis

    doi: 10.1186/s13018-021-02323-7

    Figure Lengend Snippet: Overexpression of IRAK3 partially reversed miR-591 on chondrocyte apoptosis. a Overexpression efficiency of IRAK3 was determined by western blot assay in OA chondrocytes transfected with IRAK3 or vector. b IRAK expression in OA chondrocytes transfected with NC, miR-591, miR-591 + vector, or miR-591 + IRAK3. c Cell apoptosis was determined using flow cytometry analysis. d The protein levels of Bcl-2, Bax, caspase 3, cleaved-caspase 3, caspase 9, and cleaved-caspase 9 were analyzed by western blot assay. e Western blot assay was carried out to examine the protein expression of TNF-α, IL-1β, IL-6, MMP3, and COL2A1. * P <0.05

    Article Snippet: Next, the membranes were blocked with 5% non-fat milk at room temperature for 2 h and incubated overnight at 4 °C with primary antibodies: Bcl-2 (Abcam, 1:500), Bax (Abcam, 1:500), caspase-3 (Abcam, 1:500), cleaved caspase-3 (Abcam, 1:500), caspase-9 (Abcam, 1:500), cleaved caspase-9 (Abcam, 1:500), GAPDH (Abcam, 1:500), TNF-α (Abcam, 1:500), IL-1β (Abcam, 1:500), IL-6 (Abcam, 1:500), MMP-3 (Abcam, 1:500), COL2A1 (Abcam, 1:500), and IRAK3 (Abcam, 1:500).

    Techniques: Over Expression, Western Blot, Transfection, Plasmid Preparation, Expressing, Flow Cytometry

    A) Fold changes in IRAK3 gene expression in healthy lungs after 4 hours of spontaneous breathing (non-ventilated) or mechanical ventilation with 6 ml/kg and 20 ml/kg . Bars represent the median fold-increase compared to non-ventilated animals. (*) p = 0.001 vs . non-ventilated animals. B) Representative blots from individual animals showing changes of IRAK-3 protein levels in lungs after 4 hours of mechanical ventilation with low or high V T . Histograms represent mean densitometric values showing IRAK-3 protein levels from all animals in each group (n = 6 animals per group). Data are reported as means ± SD and were obtained from 6 independent experiments. (*) p = 0.001 vs . non-ventilated animals. V T = tidal volume.

    Journal: Respiratory Research

    Article Title: Mechanical ventilation modulates TLR4 and IRAK-3 in a non-infectious, ventilator-induced lung injury model

    doi: 10.1186/1465-9921-11-27

    Figure Lengend Snippet: A) Fold changes in IRAK3 gene expression in healthy lungs after 4 hours of spontaneous breathing (non-ventilated) or mechanical ventilation with 6 ml/kg and 20 ml/kg . Bars represent the median fold-increase compared to non-ventilated animals. (*) p = 0.001 vs . non-ventilated animals. B) Representative blots from individual animals showing changes of IRAK-3 protein levels in lungs after 4 hours of mechanical ventilation with low or high V T . Histograms represent mean densitometric values showing IRAK-3 protein levels from all animals in each group (n = 6 animals per group). Data are reported as means ± SD and were obtained from 6 independent experiments. (*) p = 0.001 vs . non-ventilated animals. V T = tidal volume.

    Article Snippet: After blocking endogenous peroxidase activity (10 min in 0,3% hydrogen peroxide), slides were incubated for 1 hour at room temperature with the rabbit polyclonal anti-IRAK-3 antibody (Abcam, Cambridge, UK), then washed in PBS and incubated for 10 min with biotinylated goat anti-rabbit secondary antibody (Santa Cruz Biotechnology Inc, Santa Cruz, CA).

    Techniques: Expressing

    Immunohistochemichal localization of IRAK-3 in lung tissues of A) spontaneous breathing rats, B) rats ventilated at low (6 ml/kg) tidal volume, and C) rats ventilated at high (20 ml/kg) tidal volume . Black and white arrows in A and B point to cytoplasmatic and nuclear staining of epithelial type II cells and interstitial macrophages surrounding the alveolus, respectively. The inflammatory infiltrate with monocytes and lymphocytes and the absence of detectable IRAK-3 in C are due to the effect of high V T ventilation. Results are from at least 8 independent experiments. Tissues are counterstained with hematoxylin. Original magnification ×400.

    Journal: Respiratory Research

    Article Title: Mechanical ventilation modulates TLR4 and IRAK-3 in a non-infectious, ventilator-induced lung injury model

    doi: 10.1186/1465-9921-11-27

    Figure Lengend Snippet: Immunohistochemichal localization of IRAK-3 in lung tissues of A) spontaneous breathing rats, B) rats ventilated at low (6 ml/kg) tidal volume, and C) rats ventilated at high (20 ml/kg) tidal volume . Black and white arrows in A and B point to cytoplasmatic and nuclear staining of epithelial type II cells and interstitial macrophages surrounding the alveolus, respectively. The inflammatory infiltrate with monocytes and lymphocytes and the absence of detectable IRAK-3 in C are due to the effect of high V T ventilation. Results are from at least 8 independent experiments. Tissues are counterstained with hematoxylin. Original magnification ×400.

    Article Snippet: After blocking endogenous peroxidase activity (10 min in 0,3% hydrogen peroxide), slides were incubated for 1 hour at room temperature with the rabbit polyclonal anti-IRAK-3 antibody (Abcam, Cambridge, UK), then washed in PBS and incubated for 10 min with biotinylated goat anti-rabbit secondary antibody (Santa Cruz Biotechnology Inc, Santa Cruz, CA).

    Techniques: Staining