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ire1 activator ixa4 (MedChemExpress)

Name: ire1 activator ixa4
Brand: MedChemExpress
Rating: 94 (12 reviews)

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MedChemExpress ire1 activator ixa4

Effect of Brucella BvrR protein on <t>IRE1</t> activation and ER morphology in HMC3 cells. (A) Western blot analysis of BvrR‐His protein expression. (B) Western blot analysis of BvfA‐His protein expression. (C) Subcellular localization of BvrR‐His and ER in HMC3 cells observed by laser confocal microscopy. (D) Transmission electron microscopy analysis of ER morphology in HMC3 cells from each group. (E) Western blot analysis of p‐IRE1 protein levels. (F) Quantification of p‐IRE1 protein levels normalized to GAPDH. Data are presented as means ± standard deviations from three independent replicates; * p < 0.05 versus Control; # p < 0.05 versus pcDNA3.1; & p < 0.05 versus BvfA‐His.

Ire1 Activator Ixa4, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 12 article reviews

ire1 activator ixa4 - by Bioz Stars, 2026-02

94/100 stars

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1) Product Images from "BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB"

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

Journal: MicrobiologyOpen

doi: 10.1002/mbo3.70219

Effect of Brucella BvrR protein on IRE1 activation and ER morphology in HMC3 cells. (A) Western blot analysis of BvrR‐His protein expression. (B) Western blot analysis of BvfA‐His protein expression. (C) Subcellular localization of BvrR‐His and ER in HMC3 cells observed by laser confocal microscopy. (D) Transmission electron microscopy analysis of ER morphology in HMC3 cells from each group. (E) Western blot analysis of p‐IRE1 protein levels. (F) Quantification of p‐IRE1 protein levels normalized to GAPDH. Data are presented as means ± standard deviations from three independent replicates; * p < 0.05 versus Control; # p < 0.05 versus pcDNA3.1; & p < 0.05 versus BvfA‐His.

Figure Legend Snippet: Effect of Brucella BvrR protein on IRE1 activation and ER morphology in HMC3 cells. (A) Western blot analysis of BvrR‐His protein expression. (B) Western blot analysis of BvfA‐His protein expression. (C) Subcellular localization of BvrR‐His and ER in HMC3 cells observed by laser confocal microscopy. (D) Transmission electron microscopy analysis of ER morphology in HMC3 cells from each group. (E) Western blot analysis of p‐IRE1 protein levels. (F) Quantification of p‐IRE1 protein levels normalized to GAPDH. Data are presented as means ± standard deviations from three independent replicates; * p < 0.05 versus Control; # p < 0.05 versus pcDNA3.1; & p < 0.05 versus BvfA‐His.

Techniques Used: Activation Assay, Western Blot, Expressing, Confocal Microscopy, Transmission Assay, Electron Microscopy, Control

Brucella BvrR activates ATF2/IL‐6 and NF‐κB p65/TNF‐α signaling via IRE1. (A) Transmission electron microscopy analysis of ER morphology in HMC3 cells. (B) Western blot analysis of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α protein levels. (C–G) Quantification of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from at least three independent experiments; * p < 0.05 versus Control; # p < 0.05 versus IXA4; & p < 0.05 versus GSK2850163; ! p < 0.05 versus BvrR.

Figure Legend Snippet: Brucella BvrR activates ATF2/IL‐6 and NF‐κB p65/TNF‐α signaling via IRE1. (A) Transmission electron microscopy analysis of ER morphology in HMC3 cells. (B) Western blot analysis of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α protein levels. (C–G) Quantification of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from at least three independent experiments; * p < 0.05 versus Control; # p < 0.05 versus IXA4; & p < 0.05 versus GSK2850163; ! p < 0.05 versus BvrR.

Techniques Used: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Control

Effect of BvrR, IXA4, and GSK2850163 on IL‐6 and TNF expression. (A) ELISA measurement of IL‐6 in supernatants. (B) ELISA measurement of TNF‐α in supernatants. (C) RT‐qPCR analysis of IL6 mRNA expression. (D) RT‐qPCR analysis of TNF mRNA expression. Each assay was performed in six replicates, and data are presented as means ± standard deviations; * p < 0.05, ** p < 0.01 versus Control; # p < 0.05, ## p < 0.01 versus IXA4; & p < 0.05, && p < 0.01 versus GSK2850163; ! p < 0.05, !! p < 0.01 versus BvrR.

Figure Legend Snippet: Effect of BvrR, IXA4, and GSK2850163 on IL‐6 and TNF expression. (A) ELISA measurement of IL‐6 in supernatants. (B) ELISA measurement of TNF‐α in supernatants. (C) RT‐qPCR analysis of IL6 mRNA expression. (D) RT‐qPCR analysis of TNF mRNA expression. Each assay was performed in six replicates, and data are presented as means ± standard deviations; * p < 0.05, ** p < 0.01 versus Control; # p < 0.05, ## p < 0.01 versus IXA4; & p < 0.05, && p < 0.01 versus GSK2850163; ! p < 0.05, !! p < 0.01 versus BvrR.

Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Brucella BvrR activates IRE1, ATF2, and NF‐κB p65 in hippocampal microglia. (A) Immunofluorescence co‐localization of IBA‐1 and p‐IRE1. (B) Quantification of p‐IRE1 co‐localization with IBA‐1. (C) Immunofluorescence co‐localization of IBA‐1 and p‐ATF2. (D) Quantification of p‐ATF2 co‐localization with IBA‐1. (E) Immunofluorescence co‐localization of IBA‐1 and p‐p65. (F) Quantification of p‐p65 co‐localization with IBA‐1. (G) Western blot analysis of IL‐6 and TNF‐α protein levels in hippocampal tissue. (H) Quantification of IL‐6 expression normalized to GAPDH. (I) Quantification of TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from three independent experiments; * p < 0.05, ** p < 0.01 versus AAV‐ bvrR .

Figure Legend Snippet: Brucella BvrR activates IRE1, ATF2, and NF‐κB p65 in hippocampal microglia. (A) Immunofluorescence co‐localization of IBA‐1 and p‐IRE1. (B) Quantification of p‐IRE1 co‐localization with IBA‐1. (C) Immunofluorescence co‐localization of IBA‐1 and p‐ATF2. (D) Quantification of p‐ATF2 co‐localization with IBA‐1. (E) Immunofluorescence co‐localization of IBA‐1 and p‐p65. (F) Quantification of p‐p65 co‐localization with IBA‐1. (G) Western blot analysis of IL‐6 and TNF‐α protein levels in hippocampal tissue. (H) Quantification of IL‐6 expression normalized to GAPDH. (I) Quantification of TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from three independent experiments; * p < 0.05, ** p < 0.01 versus AAV‐ bvrR .

Techniques Used: Immunofluorescence, Western Blot, Expressing

Image Search Results

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Effect of Brucella BvrR protein on IRE1 activation and ER morphology in HMC3 cells. (A) Western blot analysis of BvrR‐His protein expression. (B) Western blot analysis of BvfA‐His protein expression. (C) Subcellular localization of BvrR‐His and ER in HMC3 cells observed by laser confocal microscopy. (D) Transmission electron microscopy analysis of ER morphology in HMC3 cells from each group. (E) Western blot analysis of p‐IRE1 protein levels. (F) Quantification of p‐IRE1 protein levels normalized to GAPDH. Data are presented as means ± standard deviations from three independent replicates; * p < 0.05 versus Control; # p < 0.05 versus pcDNA3.1; & p < 0.05 versus BvfA‐His.

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Activation Assay, Western Blot, Expressing, Confocal Microscopy, Transmission Assay, Electron Microscopy, Control

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Brucella BvrR activates ATF2/IL‐6 and NF‐κB p65/TNF‐α signaling via IRE1. (A) Transmission electron microscopy analysis of ER morphology in HMC3 cells. (B) Western blot analysis of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α protein levels. (C–G) Quantification of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from at least three independent experiments; * p < 0.05 versus Control; # p < 0.05 versus IXA4; & p < 0.05 versus GSK2850163; ! p < 0.05 versus BvrR.

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Control

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Effect of BvrR, IXA4, and GSK2850163 on IL‐6 and TNF expression. (A) ELISA measurement of IL‐6 in supernatants. (B) ELISA measurement of TNF‐α in supernatants. (C) RT‐qPCR analysis of IL6 mRNA expression. (D) RT‐qPCR analysis of TNF mRNA expression. Each assay was performed in six replicates, and data are presented as means ± standard deviations; * p < 0.05, ** p < 0.01 versus Control; # p < 0.05, ## p < 0.01 versus IXA4; & p < 0.05, && p < 0.01 versus GSK2850163; ! p < 0.05, !! p < 0.01 versus BvrR.

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Brucella BvrR activates IRE1, ATF2, and NF‐κB p65 in hippocampal microglia. (A) Immunofluorescence co‐localization of IBA‐1 and p‐IRE1. (B) Quantification of p‐IRE1 co‐localization with IBA‐1. (C) Immunofluorescence co‐localization of IBA‐1 and p‐ATF2. (D) Quantification of p‐ATF2 co‐localization with IBA‐1. (E) Immunofluorescence co‐localization of IBA‐1 and p‐p65. (F) Quantification of p‐p65 co‐localization with IBA‐1. (G) Western blot analysis of IL‐6 and TNF‐α protein levels in hippocampal tissue. (H) Quantification of IL‐6 expression normalized to GAPDH. (I) Quantification of TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from three independent experiments; * p < 0.05, ** p < 0.01 versus AAV‐ bvrR .

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Immunofluorescence, Western Blot, Expressing

( A ) Representative photomicrographs of control and XBP1 siRNA transfected myotubes incubated with or without KPC-CM. Scale bar, 50 μm. ( B ) Quantification of average myotube diameter. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( C ) Percentage reduction in average myotube diameter in cultures transfected with control or XBP1 siRNA and incubated with or without KPC-CM. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( D ) Immunoblot and ( E ) densitometry analysis of protein levels of sXBP1 in control or XBP1 siRNA transfected myotubes incubated with or without KPC-CM. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( F ) Representative photomicrographs of vehicle- or 4 μM 4μ8C-treated myotubes incubated in DM or KPC-CM are presented here. Scale bar, 50 μm. ( G ) Quantification of average myotube diameter in vehicle alone or 4μ8C-treated cultures incubated with or without KPC-CM. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( H ) Representative photomicrographs of vehicle- or 20 μM IXA4-treated myotubes incubated in DM or KPC-CM are presented here. Scale bar, 50 μm. ( I ) Quantification of average myotube diameter. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( J ) Immunoblot and ( K ) densitometry analysis of protein levels of sXBP1 in 4μ8C or IXA4-treated myotubes compared to control vehicle-treated cultures. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. .

Journal: EMBO Molecular Medicine

Article Title: The canonical ER stress IRE1α/XBP1 pathway mediates skeletal muscle wasting during pancreatic cancer cachexia

doi: 10.1038/s44321-025-00337-w

Figure Lengend Snippet: ( A ) Representative photomicrographs of control and XBP1 siRNA transfected myotubes incubated with or without KPC-CM. Scale bar, 50 μm. ( B ) Quantification of average myotube diameter. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( C ) Percentage reduction in average myotube diameter in cultures transfected with control or XBP1 siRNA and incubated with or without KPC-CM. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( D ) Immunoblot and ( E ) densitometry analysis of protein levels of sXBP1 in control or XBP1 siRNA transfected myotubes incubated with or without KPC-CM. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( F ) Representative photomicrographs of vehicle- or 4 μM 4μ8C-treated myotubes incubated in DM or KPC-CM are presented here. Scale bar, 50 μm. ( G ) Quantification of average myotube diameter in vehicle alone or 4μ8C-treated cultures incubated with or without KPC-CM. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( H ) Representative photomicrographs of vehicle- or 20 μM IXA4-treated myotubes incubated in DM or KPC-CM are presented here. Scale bar, 50 μm. ( I ) Quantification of average myotube diameter. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( J ) Immunoblot and ( K ) densitometry analysis of protein levels of sXBP1 in 4μ8C or IXA4-treated myotubes compared to control vehicle-treated cultures. n = 3 biological replicates per group. Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. .

Article Snippet: IXA4 , MedChemExpress , Cat # HY-139214.

Techniques: Control, Transfection, Incubation, Comparison, Western Blot

( A ) Gene Set Enrichment Analysis (GSEA) of DEGs shows upregulation in the gene set associated with IL6-JAK-STAT3 signaling in GA muscle of KPC tumor-bearing Xbp1 fl/fl mice compared to control Xbp1 fl/fl mice injected with PBS. ( B ) Heatmap representation shows gene expression of molecules involved in JAK-STAT signaling in GA muscle of control and KPC tumor-bearing Xbp1 fl/fl and Xbp1 mKO mice. Data information: Differentially expressed genes (DEGs) were identified using threshold of Log2FC ≥ 0.25 and P value < 0.05. ( C ) Immunoblots and ( D ) densitometry analysis showing levels of phosphorylated STAT3 (p-STAT3) and total STAT3 protein in GA muscle of control and KPC tumor-bearing Xbp1 fl/fl and Xbp1 mKO mice. n = 3–4 mice per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( E ) Immunoblots and ( F ) densitometry analysis of protein levels of p-STAT3 and STAT3 in cultures treated with vehicle alone or 20 µM IXA4. n = 3 biological replicates per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( G ) Immunoblots and ( H ) densitometry analysis of protein levels of p-STAT3 and STAT3 in myotube cultures treated with vehicle alone or 4 µM 4μ8C. n = 3 biological replicates per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( I ) Representative photomicrographs of MyHC-stained myotube cultures. Scale bar, 50 μm, ( J ) quantification of myotube diameter in vehicle- or IXA4-treated cultures transfected with control or STAT3 siRNA, ( K ) immunoblots and ( L ) densitometry analysis showing protein levels of STAT3 and sXBP1 in vehicle- or IXA4-treated cultures transfected with control or STAT3 siRNA. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. .

Journal: EMBO Molecular Medicine

Article Title: The canonical ER stress IRE1α/XBP1 pathway mediates skeletal muscle wasting during pancreatic cancer cachexia

doi: 10.1038/s44321-025-00337-w

Figure Lengend Snippet: ( A ) Gene Set Enrichment Analysis (GSEA) of DEGs shows upregulation in the gene set associated with IL6-JAK-STAT3 signaling in GA muscle of KPC tumor-bearing Xbp1 fl/fl mice compared to control Xbp1 fl/fl mice injected with PBS. ( B ) Heatmap representation shows gene expression of molecules involved in JAK-STAT signaling in GA muscle of control and KPC tumor-bearing Xbp1 fl/fl and Xbp1 mKO mice. Data information: Differentially expressed genes (DEGs) were identified using threshold of Log2FC ≥ 0.25 and P value < 0.05. ( C ) Immunoblots and ( D ) densitometry analysis showing levels of phosphorylated STAT3 (p-STAT3) and total STAT3 protein in GA muscle of control and KPC tumor-bearing Xbp1 fl/fl and Xbp1 mKO mice. n = 3–4 mice per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. ( E ) Immunoblots and ( F ) densitometry analysis of protein levels of p-STAT3 and STAT3 in cultures treated with vehicle alone or 20 µM IXA4. n = 3 biological replicates per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( G ) Immunoblots and ( H ) densitometry analysis of protein levels of p-STAT3 and STAT3 in myotube cultures treated with vehicle alone or 4 µM 4μ8C. n = 3 biological replicates per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using unpaired Student t test. ( I ) Representative photomicrographs of MyHC-stained myotube cultures. Scale bar, 50 μm, ( J ) quantification of myotube diameter in vehicle- or IXA4-treated cultures transfected with control or STAT3 siRNA, ( K ) immunoblots and ( L ) densitometry analysis showing protein levels of STAT3 and sXBP1 in vehicle- or IXA4-treated cultures transfected with control or STAT3 siRNA. n = 3 (biological replicates) per group. Data information: Data are presented as mean ± SEM. Indicated P values were calculated using two-way ANOVA followed by Tukey’s multiple comparison test. .

Article Snippet: IXA4 , MedChemExpress , Cat # HY-139214.

Techniques: Control, Injection, Gene Expression, Western Blot, Comparison, Staining, Transfection

( A , B ) Conserved promoter regions containing sXBP1 binding sequences of ( A ) CCACG-box domains upstream of Map1lc3b (LC3B) and Xbp1 genes, and ( B ) UPRE A/B domains upstream of Fbxo32 (MAFbx) and Atg5 genes are highlighted in red. ( C , D ) Chromatin immunoprecipitation (ChIP) showing enrichment of sXBP1 to the promoters of Fbxo32 , Map1lc3b , Atg5 , and Xbp1 in KPC-CM-treated myotubes analyzed by performing ( C ) semi-quantitative PCR and ( D ) quantitative PCR (qPCR). n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. ( E ) Validation of ChIP assay in KPC-CM treated cultured myotubes was performed using PCR for Histone H3 antibody binding to the promoter of Rpl30 (positive control), and sXBP1 binding to promoter regions of known targets, Dnajb9 and Hspa5 . ( F ) Conserved promoter sequences containing sXBP1 UPRE A/B domains upstream of Il6 , Pdk4 , and Stat3 genes, highlighted in red. ( G ) ChIP assay followed by PCR analysis shows enrichment of sXBP1 to the promoter regions of Fbxo32 , Xbp1 , Il6 , Pdk4 , and Stat3 genes in GA muscle of KPC-tumor bearing mice. ( H ) qPCR analysis showing fold increase in the enrichment of sXBP1 to the promoter regions of Fbxo32 , Xbp1 , Il6 , Pdk4 genes. GA muscle from 5 KPC tumor-bearing mice pooled for 1 biological sample. n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. ( I ) Validation of ChIP assay in GA muscle of tumor-bearing mice was performed using PCR for Histone H3 antibody binding to the promoter of Rpl30 (positive control) and known sXBP1 target Dnajb9 . ( J ) Relative mRNA levels of Fbxo32 , Map1lc3b , Atg5 , Il6 , Pdk4 and spliced -Xbp1 in cultured myotubes after 8 h treatment with vehicle alone or 20 μM IXA4. n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. .

Journal: EMBO Molecular Medicine

Article Title: The canonical ER stress IRE1α/XBP1 pathway mediates skeletal muscle wasting during pancreatic cancer cachexia

doi: 10.1038/s44321-025-00337-w

Figure Lengend Snippet: ( A , B ) Conserved promoter regions containing sXBP1 binding sequences of ( A ) CCACG-box domains upstream of Map1lc3b (LC3B) and Xbp1 genes, and ( B ) UPRE A/B domains upstream of Fbxo32 (MAFbx) and Atg5 genes are highlighted in red. ( C , D ) Chromatin immunoprecipitation (ChIP) showing enrichment of sXBP1 to the promoters of Fbxo32 , Map1lc3b , Atg5 , and Xbp1 in KPC-CM-treated myotubes analyzed by performing ( C ) semi-quantitative PCR and ( D ) quantitative PCR (qPCR). n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. ( E ) Validation of ChIP assay in KPC-CM treated cultured myotubes was performed using PCR for Histone H3 antibody binding to the promoter of Rpl30 (positive control), and sXBP1 binding to promoter regions of known targets, Dnajb9 and Hspa5 . ( F ) Conserved promoter sequences containing sXBP1 UPRE A/B domains upstream of Il6 , Pdk4 , and Stat3 genes, highlighted in red. ( G ) ChIP assay followed by PCR analysis shows enrichment of sXBP1 to the promoter regions of Fbxo32 , Xbp1 , Il6 , Pdk4 , and Stat3 genes in GA muscle of KPC-tumor bearing mice. ( H ) qPCR analysis showing fold increase in the enrichment of sXBP1 to the promoter regions of Fbxo32 , Xbp1 , Il6 , Pdk4 genes. GA muscle from 5 KPC tumor-bearing mice pooled for 1 biological sample. n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. ( I ) Validation of ChIP assay in GA muscle of tumor-bearing mice was performed using PCR for Histone H3 antibody binding to the promoter of Rpl30 (positive control) and known sXBP1 target Dnajb9 . ( J ) Relative mRNA levels of Fbxo32 , Map1lc3b , Atg5 , Il6 , Pdk4 and spliced -Xbp1 in cultured myotubes after 8 h treatment with vehicle alone or 20 μM IXA4. n = 3 biological replicates per group. Data information: Data are presented as mea n ± SEM. Indicated P values were calculated using unpaired Student t test. .

Article Snippet: IXA4 , MedChemExpress , Cat # HY-139214.

Techniques: Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Biomarker Discovery, Cell Culture, Positive Control

Activation of the IRE1/Xbp1s axis facilitates poly(GR) clearance in a Drosophila disease model. A , Western blot analysis showing the effects of IRE1 overexpression or IRE1-RNAi on Flag-(GR)80 protein levels in Drosophila muscle tissue. NS stands for non-specific. The bar graph quantifies relative (GR)80 levels. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001, ∗∗ p < 0.01. Genotypes are: +; MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. +; MHC>Flag-GR80/IRE1-RI . B , quantification of abnormal wing posture phenotypes induced by poly(GR) expression, with or without IRE1 overexpression or knockdown. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , Western blot analysis assessing the impact of Xbp1s overexpression or Xbp1-RNAi on Flag-(GR)80 levels in Drosophila muscle. Bar graph presents quantification of relative (GR)80 abundance. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. Genotypes are: +; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80. Xbp1-RI; MHC>Flag-GR80 . D , quantification of abnormal wing posture phenotype in flies co-expressing Flag-(GR)80 with either Xbp1s overexpression or Xbp1-RNAi. Data points are presented as the mean ± S.D.∗∗∗ p < 0.001. E , representative immunofluorescence images of Flag-tagged poly(GR) in adult Drosophila muscle upon IRE1 or Xbp1s overexpression. Scale bar: 10 μm. The corresponding bar graph shows relative Flag-poly(GR) fluorescence intensity. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. Genotypes are: +; MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80 . F , quantification of climbing ability in flies expressing Flag-(GR)80 alone or in combination with IRE1 or Xbp1s. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001, ∗ p < 0.05. G , survival analysis of flies expressing Flag-(GR)80 alone or in combination with IRE1 or Xbp1s, compared to control flies. Genotypes are: +;MHC. +;MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80 . H , Western blot analysis of Flag-(GR)80 protein levels in Drosophila muscle tissue following administration of IXA4. The bar graph displays relative poly(GR) levels. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001.

Journal: The Journal of Biological Chemistry

Article Title: IRE1/Xbp1 promotes the clearance of poly(GR) dipeptide repeats in amyotrophic lateral sclerosis

doi: 10.1016/j.jbc.2025.110764

Figure Lengend Snippet: Activation of the IRE1/Xbp1s axis facilitates poly(GR) clearance in a Drosophila disease model. A , Western blot analysis showing the effects of IRE1 overexpression or IRE1-RNAi on Flag-(GR)80 protein levels in Drosophila muscle tissue. NS stands for non-specific. The bar graph quantifies relative (GR)80 levels. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001, ∗∗ p < 0.01. Genotypes are: +; MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. +; MHC>Flag-GR80/IRE1-RI . B , quantification of abnormal wing posture phenotypes induced by poly(GR) expression, with or without IRE1 overexpression or knockdown. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , Western blot analysis assessing the impact of Xbp1s overexpression or Xbp1-RNAi on Flag-(GR)80 levels in Drosophila muscle. Bar graph presents quantification of relative (GR)80 abundance. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. Genotypes are: +; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80. Xbp1-RI; MHC>Flag-GR80 . D , quantification of abnormal wing posture phenotype in flies co-expressing Flag-(GR)80 with either Xbp1s overexpression or Xbp1-RNAi. Data points are presented as the mean ± S.D.∗∗∗ p < 0.001. E , representative immunofluorescence images of Flag-tagged poly(GR) in adult Drosophila muscle upon IRE1 or Xbp1s overexpression. Scale bar: 10 μm. The corresponding bar graph shows relative Flag-poly(GR) fluorescence intensity. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. Genotypes are: +; MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80 . F , quantification of climbing ability in flies expressing Flag-(GR)80 alone or in combination with IRE1 or Xbp1s. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001, ∗ p < 0.05. G , survival analysis of flies expressing Flag-(GR)80 alone or in combination with IRE1 or Xbp1s, compared to control flies. Genotypes are: +;MHC. +;MHC>Flag-GR80. IRE1-V5; MHC>Flag-GR80. Xbp1s-V5; MHC>Flag-GR80 . H , Western blot analysis of Flag-(GR)80 protein levels in Drosophila muscle tissue following administration of IXA4. The bar graph displays relative poly(GR) levels. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001.

Article Snippet: For Drosophila experiments, newly eclosed flies were placed in vials containing instant fly food supplemented with 200 μM IXA4 (TargetMol) and maintained for 6 days.

Techniques: Activation Assay, Western Blot, Over Expression, Expressing, Knockdown, Immunofluorescence, Fluorescence, Control

The IRE1/Xbp1s pathway supresses poly(GR) protein levels in mammalian cells and C9orf72 patient-derived fibroblasts. A , representative immunofluorescence images of Flag-tagged (GR)80 in HeLa cells following overexpression of IRE1-GFP or Xbp1s-GFP. Scale bar: 10 μm. The adjacent bar graph quantifies relative Flag-(GR)80 fluorescence intensity. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. B , Western blot analysis depicting the impact of IRE1 or Xbp1s overexpression on Flag-(GR)80 protein levels in HEK293T cells. Quantification of relative (GR)80 abundance is shown in the accompanying bar graph. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , Western blot analysis of Flag-(GR)80 protein levels in HEK293T cells treated with IXA4. The bar graph presents the relative (GR)80 levels post-treatment. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. D , Dot blot assay illustrating the effects of ectopic expression of IRE1 and Xbp1s, as well as IXA4 treatment, on endogenous poly(GR) levels in fibroblasts derived from C9orf72-ALS patients. The bar graph displays relative poly(GR) quantification. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001.

Journal: The Journal of Biological Chemistry

Article Title: IRE1/Xbp1 promotes the clearance of poly(GR) dipeptide repeats in amyotrophic lateral sclerosis

doi: 10.1016/j.jbc.2025.110764

Figure Lengend Snippet: The IRE1/Xbp1s pathway supresses poly(GR) protein levels in mammalian cells and C9orf72 patient-derived fibroblasts. A , representative immunofluorescence images of Flag-tagged (GR)80 in HeLa cells following overexpression of IRE1-GFP or Xbp1s-GFP. Scale bar: 10 μm. The adjacent bar graph quantifies relative Flag-(GR)80 fluorescence intensity. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. B , Western blot analysis depicting the impact of IRE1 or Xbp1s overexpression on Flag-(GR)80 protein levels in HEK293T cells. Quantification of relative (GR)80 abundance is shown in the accompanying bar graph. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , Western blot analysis of Flag-(GR)80 protein levels in HEK293T cells treated with IXA4. The bar graph presents the relative (GR)80 levels post-treatment. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. D , Dot blot assay illustrating the effects of ectopic expression of IRE1 and Xbp1s, as well as IXA4 treatment, on endogenous poly(GR) levels in fibroblasts derived from C9orf72-ALS patients. The bar graph displays relative poly(GR) quantification. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001.

Article Snippet: For Drosophila experiments, newly eclosed flies were placed in vials containing instant fly food supplemented with 200 μM IXA4 (TargetMol) and maintained for 6 days.

Techniques: Derivative Assay, Immunofluorescence, Over Expression, Fluorescence, Western Blot, Dot Blot, Expressing

Activation of IRE1 alleviates motor neuron degeneration in C9orf72 transgenic mice. A and B , dot blot analysis showing the effects of ectopic IRE1 expression via Lenti-IRE1 or pharmacological activation with IXA4 on poly(GR) levels in the brain and spinal cord of C9orf72 mice. Quantification of relative poly(GR) protein levels is presented in the accompanying bar graphs. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , representative immunohistochemistry (IHC) images of motor neurons stained with ChAT antibody following Lenti-IRE1 delivery or IXA4 treatment. Quantification of motor neuron counts per region is shown in the bar graph. Data points are presented as the mean ± S.D. ∗∗ p < 0.01, ∗∗∗ p < 0.001. D , representative hematoxylin and eosin (H&E) staining images of motor neurons in cortical layer V, following Lenti-IRE1 or IXA4 administration. Quantification of motor neuron numbers per region is displayed in the bar graph. Data points are presented as the mean ± S.D. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Journal: The Journal of Biological Chemistry

Article Title: IRE1/Xbp1 promotes the clearance of poly(GR) dipeptide repeats in amyotrophic lateral sclerosis

doi: 10.1016/j.jbc.2025.110764

Figure Lengend Snippet: Activation of IRE1 alleviates motor neuron degeneration in C9orf72 transgenic mice. A and B , dot blot analysis showing the effects of ectopic IRE1 expression via Lenti-IRE1 or pharmacological activation with IXA4 on poly(GR) levels in the brain and spinal cord of C9orf72 mice. Quantification of relative poly(GR) protein levels is presented in the accompanying bar graphs. Data points are presented as the mean ± S.D. ∗∗∗ p < 0.001. C , representative immunohistochemistry (IHC) images of motor neurons stained with ChAT antibody following Lenti-IRE1 delivery or IXA4 treatment. Quantification of motor neuron counts per region is shown in the bar graph. Data points are presented as the mean ± S.D. ∗∗ p < 0.01, ∗∗∗ p < 0.001. D , representative hematoxylin and eosin (H&E) staining images of motor neurons in cortical layer V, following Lenti-IRE1 or IXA4 administration. Quantification of motor neuron numbers per region is displayed in the bar graph. Data points are presented as the mean ± S.D. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: For Drosophila experiments, newly eclosed flies were placed in vials containing instant fly food supplemented with 200 μM IXA4 (TargetMol) and maintained for 6 days.

Techniques: Activation Assay, Transgenic Assay, Dot Blot, Expressing, Immunohistochemistry, Staining

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1) Product Images from "BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB"

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