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sh redd1  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology sh redd1
    a Weight gain in <t>Redd1</t> −/− mice and their WT littermates fed NC or HFD for 16 weeks ( n = 6 per group). b Mass of eWAT and iWAT in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 8 per group). c Representative images of perilipin (green) and F4/80 (purple) staining in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). Scale bar, 100 μm. d Average adipocyte size in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). e Relative area of F4/80-positive cells in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Relative number of crown-like structures (CLSs) in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). g NF-κB activity in the eWAT from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). h Plasma levels of inflammatory cytokines in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.
    Sh Redd1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sh redd1/product/Santa Cruz Biotechnology
    Average 93 stars, based on 2 article reviews
    sh redd1 - by Bioz Stars, 2026-03
    93/100 stars

    Images

    1) Product Images from "REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation"

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    Journal: Nature Communications

    doi: 10.1038/s41467-022-34110-1

    a Weight gain in Redd1 −/− mice and their WT littermates fed NC or HFD for 16 weeks ( n = 6 per group). b Mass of eWAT and iWAT in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 8 per group). c Representative images of perilipin (green) and F4/80 (purple) staining in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). Scale bar, 100 μm. d Average adipocyte size in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). e Relative area of F4/80-positive cells in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Relative number of crown-like structures (CLSs) in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). g NF-κB activity in the eWAT from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). h Plasma levels of inflammatory cytokines in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Weight gain in Redd1 −/− mice and their WT littermates fed NC or HFD for 16 weeks ( n = 6 per group). b Mass of eWAT and iWAT in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 8 per group). c Representative images of perilipin (green) and F4/80 (purple) staining in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). Scale bar, 100 μm. d Average adipocyte size in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). e Relative area of F4/80-positive cells in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Relative number of crown-like structures (CLSs) in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). g NF-κB activity in the eWAT from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). h Plasma levels of inflammatory cytokines in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Techniques Used: Staining, Activity Assay, Clinical Proteomics

    a Fasting plasma levels of glucose and insulin in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 6 per group). b Representative images of insulin (green)-stained pancreatic islets from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Scale bar, 100 μm. c Quantification of average islet size ( n = 6 per group). d Calculation of the HOMA-IR scores ( n = 6 per group). e Assessment of GTT and ITT in mice fasting for 12 and 6 h, respectively, in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Representative western blots of Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) in eWAT and skeletal muscle from mice injected i.p. with saline or insulin ( n = 3). g Representative western blots of phosphorylated Akt and FOXO1 in the liver of mice injected with saline or insulin ( n = 6). h Quantification of the phosphorylated FOXO1 to total FOXO1 ratio ( n = 6 per group). i Quantification of G6pc , Pck1 , and Fbp1 mRNA levels in the liver ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Fasting plasma levels of glucose and insulin in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 6 per group). b Representative images of insulin (green)-stained pancreatic islets from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Scale bar, 100 μm. c Quantification of average islet size ( n = 6 per group). d Calculation of the HOMA-IR scores ( n = 6 per group). e Assessment of GTT and ITT in mice fasting for 12 and 6 h, respectively, in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Representative western blots of Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) in eWAT and skeletal muscle from mice injected i.p. with saline or insulin ( n = 3). g Representative western blots of phosphorylated Akt and FOXO1 in the liver of mice injected with saline or insulin ( n = 6). h Quantification of the phosphorylated FOXO1 to total FOXO1 ratio ( n = 6 per group). i Quantification of G6pc , Pck1 , and Fbp1 mRNA levels in the liver ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Techniques Used: Clinical Proteomics, Staining, Western Blot, Phospho-proteomics, Membrane, Injection, Saline

    a Weight gain over time in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) mice fed HFD for 16 weeks ( n = 6 per group). b Mass measurements for the eWAT and iWAT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of Redd1 fl/fl and Redd1 Δ Adipoq mice fed HFD ( n = 6 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). e , f Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). g Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of phosphorylated IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylated Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after i.p. injection of saline or insulin ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice compared with NC-fed mouse groups ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t-test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Weight gain over time in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) mice fed HFD for 16 weeks ( n = 6 per group). b Mass measurements for the eWAT and iWAT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of Redd1 fl/fl and Redd1 Δ Adipoq mice fed HFD ( n = 6 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). e , f Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). g Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of phosphorylated IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylated Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after i.p. injection of saline or insulin ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice compared with NC-fed mouse groups ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t-test. Source data are provided as a Source Data file.

    Techniques Used: Staining, Activity Assay, Clinical Proteomics, Western Blot, Injection, Saline, Expressing, Two Tailed Test

    a Weight gain in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ LysM ( R Δ LysM ) mice fed HFD for 16 weeks ( n = 5 per group). b Measurement of fat (eWAT + iWAT) mass in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). e Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). f Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). g Calculation of HOMA-IR scores in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of the insulin-responsive phosphorylation of IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylation of Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice compared with NC-fed mouse groups ( n = 5 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Weight gain in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ LysM ( R Δ LysM ) mice fed HFD for 16 weeks ( n = 5 per group). b Measurement of fat (eWAT + iWAT) mass in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). e Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). f Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). g Calculation of HOMA-IR scores in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of the insulin-responsive phosphorylation of IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylation of Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice compared with NC-fed mouse groups ( n = 5 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Techniques Used: Staining, Activity Assay, Clinical Proteomics, Western Blot, Phospho-proteomics, Expressing, Two Tailed Test

    a – c Representative oil red-O (ORO)-stained images of WT and Redd1 −/− SVF cells ( a ), shControl (shC)- or sh-Redd1-transfected 3T3-L1 cells ( b ), and WT ( Redd1 fl/f l , R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) SVF cells ( c ) when cultured in differentiation medium (MDI) and quantification of relative ORO intensity ( n = 4). d – f , Expression levels of adipogenic genes ( d ), REDD1 ( e ), and lipogenic genes ( f ) in R fl/fl and R Δ Adipoq SVF cells cultured in MDI medium and quantification of relative ORO intensity ( n = 4). g Assessment of NF-κB–Luc activity in 3T3-L1 cells transfected either with siRNA for control, Ikka , Ikkb , or NF-κB p65 ( p65 ) or with pcDNA3.1/His- Ikba ( n = 5). h , i Representative images and realative quantification of ORO-stained images ( h ) and expression levels of Pparg and Cebpa ( i ) in 3T3-L1 cells infected with control adenovirus (Ad-C) or adenoviral Redd1 (Ad- R ) after transfection with vector alone or pcDNA3.1/His- Ikba ( n = 4). j NF-κB–Luc activity in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 4). k Cytokine production in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 5). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( g , h ) and two-way ANOVA ( a , i ) followed by the Holm–Sidak post hoc test and an unpaired two-tailed t -test ( b – f , j , k ). Source data are provided as a Source Data file.
    Figure Legend Snippet: a – c Representative oil red-O (ORO)-stained images of WT and Redd1 −/− SVF cells ( a ), shControl (shC)- or sh-Redd1-transfected 3T3-L1 cells ( b ), and WT ( Redd1 fl/f l , R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) SVF cells ( c ) when cultured in differentiation medium (MDI) and quantification of relative ORO intensity ( n = 4). d – f , Expression levels of adipogenic genes ( d ), REDD1 ( e ), and lipogenic genes ( f ) in R fl/fl and R Δ Adipoq SVF cells cultured in MDI medium and quantification of relative ORO intensity ( n = 4). g Assessment of NF-κB–Luc activity in 3T3-L1 cells transfected either with siRNA for control, Ikka , Ikkb , or NF-κB p65 ( p65 ) or with pcDNA3.1/His- Ikba ( n = 5). h , i Representative images and realative quantification of ORO-stained images ( h ) and expression levels of Pparg and Cebpa ( i ) in 3T3-L1 cells infected with control adenovirus (Ad-C) or adenoviral Redd1 (Ad- R ) after transfection with vector alone or pcDNA3.1/His- Ikba ( n = 4). j NF-κB–Luc activity in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 4). k Cytokine production in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 5). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( g , h ) and two-way ANOVA ( a , i ) followed by the Holm–Sidak post hoc test and an unpaired two-tailed t -test ( b – f , j , k ). Source data are provided as a Source Data file.

    Techniques Used: Staining, Transfection, Cell Culture, Expressing, Activity Assay, Control, Infection, Plasmid Preparation, Two Tailed Test

    a Predictive binding conformation between REDD1 and IκBα using computational protein-protein molecular docking methods. b Co-immunoprecipitation analysis of the interaction between REDD1 and IκBα in HEK293 cells transfected with pcDNA3.1/His- Ikba (His- Ikba ) and either pFlag-CMV-1- Redd1 ( Redd1 ) or Redd1 mutants ( R KKAA and R KKRAAA ) ( n = 3). c Representative confocal images of NF-κB p65 nuclear translocalization in HEK293 cells infected with Ad-C, Ad- Redd1 , or its mutants ( n = 4). Scale bar, 50 μm. d Assessment of NF-κB–Luc activity in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or its mutants ( n = 4). e Representative ORO-stained images of 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA and quantification of relative ORO intensity ( n = 4). f Expression levels of Pparg and Cebpa in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). g Production of MCP-1 and TNF-α in macrophages infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( d , e ) and two-way ANOVA ( f , g ) followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Predictive binding conformation between REDD1 and IκBα using computational protein-protein molecular docking methods. b Co-immunoprecipitation analysis of the interaction between REDD1 and IκBα in HEK293 cells transfected with pcDNA3.1/His- Ikba (His- Ikba ) and either pFlag-CMV-1- Redd1 ( Redd1 ) or Redd1 mutants ( R KKAA and R KKRAAA ) ( n = 3). c Representative confocal images of NF-κB p65 nuclear translocalization in HEK293 cells infected with Ad-C, Ad- Redd1 , or its mutants ( n = 4). Scale bar, 50 μm. d Assessment of NF-κB–Luc activity in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or its mutants ( n = 4). e Representative ORO-stained images of 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA and quantification of relative ORO intensity ( n = 4). f Expression levels of Pparg and Cebpa in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). g Production of MCP-1 and TNF-α in macrophages infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( d , e ) and two-way ANOVA ( f , g ) followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Techniques Used: Binding Assay, Immunoprecipitation, Transfection, Infection, Activity Assay, Control, Staining, Expressing

    a Weight gain in WT and Redd1 KKAA mice after being fed HFD for 16 weeks ( n = 10 per group). b eWAT and iWAT mass measurements in HFD-fed Redd1 KKAA mice and their WT littermates ( n = 10 per group). c Expression levels of Pparg and Cebpa in the eWAT of Redd1 KKAA mice and WT littermates fed HFD for 10 weeks ( n = 8 per group). d Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 KKAA mice and WT littermates ( n = 5 per group). Scale bar, 100 μm. e NF-κB activity in the eWAT from HFD-fed Redd1 KKAA mice and WT littermates ( n = 6 per group). f , g Plasma levels of inflammatory cytokines in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). h Fasting plasma levels of glucose and insulin in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). i Assessment of GTT and ITT in HFD-fed Redd1 KKAA mice and WT littermates after fasting for 12 and 6 h, respectively ( n = 6 per group). j , k Representative western blots of insulin-responsive Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) levels in the eWAT and skeletal muscle ( j ) and Akt and FOXO1 phosphorylation in the liver ( k ) of HFD-fed Redd1 KKAA mice and WT littermates ( n = 3). l Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 KKAA mice and WT littermates compared with NC-fed mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Weight gain in WT and Redd1 KKAA mice after being fed HFD for 16 weeks ( n = 10 per group). b eWAT and iWAT mass measurements in HFD-fed Redd1 KKAA mice and their WT littermates ( n = 10 per group). c Expression levels of Pparg and Cebpa in the eWAT of Redd1 KKAA mice and WT littermates fed HFD for 10 weeks ( n = 8 per group). d Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 KKAA mice and WT littermates ( n = 5 per group). Scale bar, 100 μm. e NF-κB activity in the eWAT from HFD-fed Redd1 KKAA mice and WT littermates ( n = 6 per group). f , g Plasma levels of inflammatory cytokines in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). h Fasting plasma levels of glucose and insulin in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). i Assessment of GTT and ITT in HFD-fed Redd1 KKAA mice and WT littermates after fasting for 12 and 6 h, respectively ( n = 6 per group). j , k Representative western blots of insulin-responsive Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) levels in the eWAT and skeletal muscle ( j ) and Akt and FOXO1 phosphorylation in the liver ( k ) of HFD-fed Redd1 KKAA mice and WT littermates ( n = 3). l Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 KKAA mice and WT littermates compared with NC-fed mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Techniques Used: Expressing, Staining, Activity Assay, Clinical Proteomics, Western Blot, Phospho-proteomics, Membrane, Two Tailed Test

    a Representative images of H&E-stained liver tissues from HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and control mice, and quantification of hepatic steatosis from H&E-stained liver tissues ( n = 6 per group). Scale bars, 100 μm. b Expression levels of Acc , Fasn , and Scd-1 in the liver of HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and their control mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Representative images of H&E-stained liver tissues from HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and control mice, and quantification of hepatic steatosis from H&E-stained liver tissues ( n = 6 per group). Scale bars, 100 μm. b Expression levels of Acc , Fasn , and Scd-1 in the liver of HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and their control mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Techniques Used: Staining, Control, Expressing, Two Tailed Test



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    99
    MedChemExpress sh redd1 dmso
    NFIL3 promotes the transcription of <t>REDD1.</t> (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.
    Sh Redd1 Dmso, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    redd1 shrna  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology redd1 shrna
    NFIL3 promotes the transcription of <t>REDD1.</t> (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.
    Redd1 Shrna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    NFIL3 promotes the transcription of REDD1. (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 promotes the transcription of REDD1. (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Assay, Over Expression, Standard Deviation

    REDD1 inactivates the mTOR pathway to trigger neutrophil autophagy and inflammatory response. (A) REDD1-related genes analyzed by the GeneMANIA database. Each circle in the figure represents a gene, and the line between circles indicates interaction between two genes. (B) KEGG enrichment analysis of REDD1-related genes. The abscissa represents GeneRatio, the ordinate represents the entry identifier, the color represents enrichment p -value, and the size represents the number of enriched genes in the identifier. (C) Western blot analysis of p-S6 and p-4EBP in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. (D) Levels of IL-17, IL-23 and IL-1β in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. (E) Western blot analysis of p-S6 and p-4EBP in the MSU-treated neutrophils. (F) Levels of IL-17, IL-23 and IL-1β in the MSU-treated cell supernatant measured by ELISA. (G) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1. (H) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1. (I) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1 or combined with Rapamycin. (J) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1 or combined with Rapamycin. (K) Levels of IL-17, IL-23 and IL-1β in the supernatant of cells treated with sh-REDD1 or combined with Rapamycin measured by ELISA. In (C,D) , *** p < 0.001, compared to healthy controls. In (E,F) , ** p < 0.01, compared to cells treated with PBS; *** p < 0.001, compared to cells treated with PBS. In (G,H) , ** p < 0.01, compared to cells treated with sh-NC; *** p < 0.001, compared to cells treated with sh-NC. In (I–K) , ** p < 0.01, compared to cells treated with sh-NC + DMSO; ## p < 0.01, compared to cells treated with sh-REDD1 + DMSO; ### p < 0.001, compared to cells treated with sh-REDD1 + DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: REDD1 inactivates the mTOR pathway to trigger neutrophil autophagy and inflammatory response. (A) REDD1-related genes analyzed by the GeneMANIA database. Each circle in the figure represents a gene, and the line between circles indicates interaction between two genes. (B) KEGG enrichment analysis of REDD1-related genes. The abscissa represents GeneRatio, the ordinate represents the entry identifier, the color represents enrichment p -value, and the size represents the number of enriched genes in the identifier. (C) Western blot analysis of p-S6 and p-4EBP in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. (D) Levels of IL-17, IL-23 and IL-1β in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. (E) Western blot analysis of p-S6 and p-4EBP in the MSU-treated neutrophils. (F) Levels of IL-17, IL-23 and IL-1β in the MSU-treated cell supernatant measured by ELISA. (G) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1. (H) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1. (I) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1 or combined with Rapamycin. (J) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1 or combined with Rapamycin. (K) Levels of IL-17, IL-23 and IL-1β in the supernatant of cells treated with sh-REDD1 or combined with Rapamycin measured by ELISA. In (C,D) , *** p < 0.001, compared to healthy controls. In (E,F) , ** p < 0.01, compared to cells treated with PBS; *** p < 0.001, compared to cells treated with PBS. In (G,H) , ** p < 0.01, compared to cells treated with sh-NC; *** p < 0.001, compared to cells treated with sh-NC. In (I–K) , ** p < 0.01, compared to cells treated with sh-NC + DMSO; ## p < 0.01, compared to cells treated with sh-REDD1 + DMSO; ### p < 0.001, compared to cells treated with sh-REDD1 + DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Standard Deviation

    REDD1 induces the formation of NETs by promoting neutrophil autophagy. (A) Levels of NETs in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. *** p < 0.001, compared to healthy controls. (B) Immunofluorescence staining analysis of the formation of NETs in MSU-treated neutrophils, scale bar = 50 μm. Neutrophil elastase (NE) and DAPI (DNA) were used to stain the cells. The white arrow represents the formed NETs. *** p < 0.001, compared to PBS-treated neutrophils. (C) Western blot analysis of LC3 II protein in MSU-induced neutrophils treated with Baf A1. ** p < 0.001, compared to MSU-treated neutrophils. (D) Immunofluorescence staining analysis of the formation of NETs in MSU-induced neutrophils treated with Baf A1. ** p < 0.01, compared to MSU-treated neutrophils. (E) Immunofluorescence staining analysis of the formation of NETs in neutrophils treated with sh-REDD1 or combined with Rapamycin, scale bar = 50 μm. ** p < 0.01, compared to neutrophils treated with sh-NC and DMSO. ## p < 0.01, compared to neutrophils treated with sh-REDD1 and DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: REDD1 induces the formation of NETs by promoting neutrophil autophagy. (A) Levels of NETs in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. *** p < 0.001, compared to healthy controls. (B) Immunofluorescence staining analysis of the formation of NETs in MSU-treated neutrophils, scale bar = 50 μm. Neutrophil elastase (NE) and DAPI (DNA) were used to stain the cells. The white arrow represents the formed NETs. *** p < 0.001, compared to PBS-treated neutrophils. (C) Western blot analysis of LC3 II protein in MSU-induced neutrophils treated with Baf A1. ** p < 0.001, compared to MSU-treated neutrophils. (D) Immunofluorescence staining analysis of the formation of NETs in MSU-induced neutrophils treated with Baf A1. ** p < 0.01, compared to MSU-treated neutrophils. (E) Immunofluorescence staining analysis of the formation of NETs in neutrophils treated with sh-REDD1 or combined with Rapamycin, scale bar = 50 μm. ** p < 0.01, compared to neutrophils treated with sh-NC and DMSO. ## p < 0.01, compared to neutrophils treated with sh-REDD1 and DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Western Blot, Standard Deviation

    NFIL3 silencing contributes to suppressed neutrophil autophagy and NET formation, which is related to downregulated expression of REDD1. (A) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (B) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (C) Western blot analysis of p-S6 and p-4EBP1 in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (D) Western blot analysis of LC3 II protein in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (E) NET formation in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1, scale bar = 50 μm. *** p < 0.001, compared to neutrophils treated with sh-NC + Ad-NC. ## p < 0.01, compared to neutrophils treated with sh-NFIL3 + Ad-NC. ### p < 0.001, compared to neutrophils treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 silencing contributes to suppressed neutrophil autophagy and NET formation, which is related to downregulated expression of REDD1. (A) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (B) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (C) Western blot analysis of p-S6 and p-4EBP1 in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (D) Western blot analysis of LC3 II protein in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (E) NET formation in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1, scale bar = 50 μm. *** p < 0.001, compared to neutrophils treated with sh-NC + Ad-NC. ## p < 0.01, compared to neutrophils treated with sh-NFIL3 + Ad-NC. ### p < 0.001, compared to neutrophils treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

    NFIL3 silencing impairs neutrophil autophagy, reduces NET formation and inflammatory injury in mice with acute gouty arthritis via the REDD1/mTOR axis. PBS-treated mice served as the control and MSU mice were treated with sh-NFIL3 or combined with Ad-REDD1. (A) Representative microscopic views of ankle joint swelling of mice (the left), and the corresponding quantitative analysis (the right). (B) The representative images of pathological changes in the joint tissues of mice following HE staining, scale bar = 25 μm. (C) mRNA expression of NFIL3 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (D) mRNA expression of REDD1 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (E) Western blot analysis of NFIL3 expression in the neutrophils of the joint tissues of mice. (F) Quantitative analysis of REDD1 protein expression. (G) Quantitative analysis of S6 and 4EBP phosphorylation levels. (H) Quantitative analysis of LC3 II protein expression. (I) MPO activity in the joint tissue of mice. (J) IL-17 levels measured by ELISA in the serum of mice. (K) IL-23 levels measured by ELISA in the serum of mice. (L) IL-1β levels measured by ELISA in the serum of mice. (M) Quantitative analysis of NET formation in the joint tissue of mice. *** p < 0.001, compared to PBS-treated mice. ## p < 0.01, compared to MSU mice treated with sh-NC + Ad-NC. ### p < 0.001, compared to MSU mice treated with sh-NC + Ad-NC. &, && or &&& p < 0.05, compared to MSU mice treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA. n = 10 for mice in each group.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 silencing impairs neutrophil autophagy, reduces NET formation and inflammatory injury in mice with acute gouty arthritis via the REDD1/mTOR axis. PBS-treated mice served as the control and MSU mice were treated with sh-NFIL3 or combined with Ad-REDD1. (A) Representative microscopic views of ankle joint swelling of mice (the left), and the corresponding quantitative analysis (the right). (B) The representative images of pathological changes in the joint tissues of mice following HE staining, scale bar = 25 μm. (C) mRNA expression of NFIL3 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (D) mRNA expression of REDD1 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (E) Western blot analysis of NFIL3 expression in the neutrophils of the joint tissues of mice. (F) Quantitative analysis of REDD1 protein expression. (G) Quantitative analysis of S6 and 4EBP phosphorylation levels. (H) Quantitative analysis of LC3 II protein expression. (I) MPO activity in the joint tissue of mice. (J) IL-17 levels measured by ELISA in the serum of mice. (K) IL-23 levels measured by ELISA in the serum of mice. (L) IL-1β levels measured by ELISA in the serum of mice. (M) Quantitative analysis of NET formation in the joint tissue of mice. *** p < 0.001, compared to PBS-treated mice. ## p < 0.01, compared to MSU mice treated with sh-NC + Ad-NC. ### p < 0.001, compared to MSU mice treated with sh-NC + Ad-NC. &, && or &&& p < 0.05, compared to MSU mice treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA. n = 10 for mice in each group.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Control, Staining, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Schematic diagram of the mechanism by which NFIL3 affects gout. NFIL3 promotes the transcription of REDD1 and inhibits the mTOR pathway, thus activating the autophagy of neutrophils and inducing the formation of NETs, eventually aggravating the inflammatory response of gout.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: Schematic diagram of the mechanism by which NFIL3 affects gout. NFIL3 promotes the transcription of REDD1 and inhibits the mTOR pathway, thus activating the autophagy of neutrophils and inducing the formation of NETs, eventually aggravating the inflammatory response of gout.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques:

    a Weight gain in Redd1 −/− mice and their WT littermates fed NC or HFD for 16 weeks ( n = 6 per group). b Mass of eWAT and iWAT in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 8 per group). c Representative images of perilipin (green) and F4/80 (purple) staining in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). Scale bar, 100 μm. d Average adipocyte size in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). e Relative area of F4/80-positive cells in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Relative number of crown-like structures (CLSs) in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). g NF-κB activity in the eWAT from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). h Plasma levels of inflammatory cytokines in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Weight gain in Redd1 −/− mice and their WT littermates fed NC or HFD for 16 weeks ( n = 6 per group). b Mass of eWAT and iWAT in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 8 per group). c Representative images of perilipin (green) and F4/80 (purple) staining in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). Scale bar, 100 μm. d Average adipocyte size in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). e Relative area of F4/80-positive cells in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Relative number of crown-like structures (CLSs) in the eWAT of NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). g NF-κB activity in the eWAT from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). h Plasma levels of inflammatory cytokines in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Staining, Activity Assay, Clinical Proteomics

    a Fasting plasma levels of glucose and insulin in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 6 per group). b Representative images of insulin (green)-stained pancreatic islets from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Scale bar, 100 μm. c Quantification of average islet size ( n = 6 per group). d Calculation of the HOMA-IR scores ( n = 6 per group). e Assessment of GTT and ITT in mice fasting for 12 and 6 h, respectively, in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Representative western blots of Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) in eWAT and skeletal muscle from mice injected i.p. with saline or insulin ( n = 3). g Representative western blots of phosphorylated Akt and FOXO1 in the liver of mice injected with saline or insulin ( n = 6). h Quantification of the phosphorylated FOXO1 to total FOXO1 ratio ( n = 6 per group). i Quantification of G6pc , Pck1 , and Fbp1 mRNA levels in the liver ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Fasting plasma levels of glucose and insulin in NC- or HFD-fed Redd1 −/− mice and their WT littermates ( n = 6 per group). b Representative images of insulin (green)-stained pancreatic islets from NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 6 per group). Scale bar, 100 μm. c Quantification of average islet size ( n = 6 per group). d Calculation of the HOMA-IR scores ( n = 6 per group). e Assessment of GTT and ITT in mice fasting for 12 and 6 h, respectively, in NC- or HFD-fed Redd1 −/− mice and WT littermates ( n = 8 per group). f Representative western blots of Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) in eWAT and skeletal muscle from mice injected i.p. with saline or insulin ( n = 3). g Representative western blots of phosphorylated Akt and FOXO1 in the liver of mice injected with saline or insulin ( n = 6). h Quantification of the phosphorylated FOXO1 to total FOXO1 ratio ( n = 6 per group). i Quantification of G6pc , Pck1 , and Fbp1 mRNA levels in the liver ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using two-way ANOVA followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Clinical Proteomics, Staining, Western Blot, Phospho-proteomics, Membrane, Injection, Saline

    a Weight gain over time in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) mice fed HFD for 16 weeks ( n = 6 per group). b Mass measurements for the eWAT and iWAT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of Redd1 fl/fl and Redd1 Δ Adipoq mice fed HFD ( n = 6 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). e , f Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). g Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of phosphorylated IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylated Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after i.p. injection of saline or insulin ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice compared with NC-fed mouse groups ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t-test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Weight gain over time in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) mice fed HFD for 16 weeks ( n = 6 per group). b Mass measurements for the eWAT and iWAT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of Redd1 fl/fl and Redd1 Δ Adipoq mice fed HFD ( n = 6 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). e , f Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). g Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice ( n = 6 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of phosphorylated IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylated Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice after i.p. injection of saline or insulin ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ Adipoq mice compared with NC-fed mouse groups ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t-test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Staining, Activity Assay, Clinical Proteomics, Western Blot, Injection, Saline, Expressing, Two Tailed Test

    a Weight gain in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ LysM ( R Δ LysM ) mice fed HFD for 16 weeks ( n = 5 per group). b Measurement of fat (eWAT + iWAT) mass in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). e Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). f Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). g Calculation of HOMA-IR scores in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of the insulin-responsive phosphorylation of IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylation of Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice compared with NC-fed mouse groups ( n = 5 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Weight gain in Redd1 fl/fl ( R fl/fl ) and Redd1 Δ LysM ( R Δ LysM ) mice fed HFD for 16 weeks ( n = 5 per group). b Measurement of fat (eWAT + iWAT) mass in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). c Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). Scale bar, 100 μm. d NF-κB activity in the eWAT from HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). e Plasma levels of inflammatory cytokines in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). f Fasting plasma levels of glucose and insulin in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). g Calculation of HOMA-IR scores in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 5 per group). h Assessment of GTT and ITT in HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice fasting for 12 and 6 h, respectively ( n = 5 per group). i , j Representative western blots of the insulin-responsive phosphorylation of IRS-1 and Akt in the eWAT and skeletal muscle ( i ) and phosphorylation of Akt and FOXO1 in the liver ( j ) of NC- or HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice ( n = 3). k Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 fl/fl and Redd1 Δ LysM mice compared with NC-fed mouse groups ( n = 5 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Staining, Activity Assay, Clinical Proteomics, Western Blot, Phospho-proteomics, Expressing, Two Tailed Test

    a – c Representative oil red-O (ORO)-stained images of WT and Redd1 −/− SVF cells ( a ), shControl (shC)- or sh-Redd1-transfected 3T3-L1 cells ( b ), and WT ( Redd1 fl/f l , R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) SVF cells ( c ) when cultured in differentiation medium (MDI) and quantification of relative ORO intensity ( n = 4). d – f , Expression levels of adipogenic genes ( d ), REDD1 ( e ), and lipogenic genes ( f ) in R fl/fl and R Δ Adipoq SVF cells cultured in MDI medium and quantification of relative ORO intensity ( n = 4). g Assessment of NF-κB–Luc activity in 3T3-L1 cells transfected either with siRNA for control, Ikka , Ikkb , or NF-κB p65 ( p65 ) or with pcDNA3.1/His- Ikba ( n = 5). h , i Representative images and realative quantification of ORO-stained images ( h ) and expression levels of Pparg and Cebpa ( i ) in 3T3-L1 cells infected with control adenovirus (Ad-C) or adenoviral Redd1 (Ad- R ) after transfection with vector alone or pcDNA3.1/His- Ikba ( n = 4). j NF-κB–Luc activity in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 4). k Cytokine production in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 5). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( g , h ) and two-way ANOVA ( a , i ) followed by the Holm–Sidak post hoc test and an unpaired two-tailed t -test ( b – f , j , k ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a – c Representative oil red-O (ORO)-stained images of WT and Redd1 −/− SVF cells ( a ), shControl (shC)- or sh-Redd1-transfected 3T3-L1 cells ( b ), and WT ( Redd1 fl/f l , R fl/fl ) and Redd1 Δ Adipoq ( R Δ Adipoq ) SVF cells ( c ) when cultured in differentiation medium (MDI) and quantification of relative ORO intensity ( n = 4). d – f , Expression levels of adipogenic genes ( d ), REDD1 ( e ), and lipogenic genes ( f ) in R fl/fl and R Δ Adipoq SVF cells cultured in MDI medium and quantification of relative ORO intensity ( n = 4). g Assessment of NF-κB–Luc activity in 3T3-L1 cells transfected either with siRNA for control, Ikka , Ikkb , or NF-κB p65 ( p65 ) or with pcDNA3.1/His- Ikba ( n = 5). h , i Representative images and realative quantification of ORO-stained images ( h ) and expression levels of Pparg and Cebpa ( i ) in 3T3-L1 cells infected with control adenovirus (Ad-C) or adenoviral Redd1 (Ad- R ) after transfection with vector alone or pcDNA3.1/His- Ikba ( n = 4). j NF-κB–Luc activity in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 4). k Cytokine production in mouse peritoneal macrophages infected with Ad-C or Ad- Redd1 ( n = 5). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( g , h ) and two-way ANOVA ( a , i ) followed by the Holm–Sidak post hoc test and an unpaired two-tailed t -test ( b – f , j , k ). Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Staining, Transfection, Cell Culture, Expressing, Activity Assay, Control, Infection, Plasmid Preparation, Two Tailed Test

    a Predictive binding conformation between REDD1 and IκBα using computational protein-protein molecular docking methods. b Co-immunoprecipitation analysis of the interaction between REDD1 and IκBα in HEK293 cells transfected with pcDNA3.1/His- Ikba (His- Ikba ) and either pFlag-CMV-1- Redd1 ( Redd1 ) or Redd1 mutants ( R KKAA and R KKRAAA ) ( n = 3). c Representative confocal images of NF-κB p65 nuclear translocalization in HEK293 cells infected with Ad-C, Ad- Redd1 , or its mutants ( n = 4). Scale bar, 50 μm. d Assessment of NF-κB–Luc activity in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or its mutants ( n = 4). e Representative ORO-stained images of 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA and quantification of relative ORO intensity ( n = 4). f Expression levels of Pparg and Cebpa in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). g Production of MCP-1 and TNF-α in macrophages infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( d , e ) and two-way ANOVA ( f , g ) followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Predictive binding conformation between REDD1 and IκBα using computational protein-protein molecular docking methods. b Co-immunoprecipitation analysis of the interaction between REDD1 and IκBα in HEK293 cells transfected with pcDNA3.1/His- Ikba (His- Ikba ) and either pFlag-CMV-1- Redd1 ( Redd1 ) or Redd1 mutants ( R KKAA and R KKRAAA ) ( n = 3). c Representative confocal images of NF-κB p65 nuclear translocalization in HEK293 cells infected with Ad-C, Ad- Redd1 , or its mutants ( n = 4). Scale bar, 50 μm. d Assessment of NF-κB–Luc activity in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or its mutants ( n = 4). e Representative ORO-stained images of 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA and quantification of relative ORO intensity ( n = 4). f Expression levels of Pparg and Cebpa in 3T3-L1 cells infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). g Production of MCP-1 and TNF-α in macrophages infected with Ad-control, Ad- Redd1 , or Ad- Redd1 KKAA ( n = 4). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using one-way ANOVA ( d , e ) and two-way ANOVA ( f , g ) followed by the Holm–Sidak post hoc test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Binding Assay, Immunoprecipitation, Transfection, Infection, Activity Assay, Control, Staining, Expressing

    a Weight gain in WT and Redd1 KKAA mice after being fed HFD for 16 weeks ( n = 10 per group). b eWAT and iWAT mass measurements in HFD-fed Redd1 KKAA mice and their WT littermates ( n = 10 per group). c Expression levels of Pparg and Cebpa in the eWAT of Redd1 KKAA mice and WT littermates fed HFD for 10 weeks ( n = 8 per group). d Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 KKAA mice and WT littermates ( n = 5 per group). Scale bar, 100 μm. e NF-κB activity in the eWAT from HFD-fed Redd1 KKAA mice and WT littermates ( n = 6 per group). f , g Plasma levels of inflammatory cytokines in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). h Fasting plasma levels of glucose and insulin in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). i Assessment of GTT and ITT in HFD-fed Redd1 KKAA mice and WT littermates after fasting for 12 and 6 h, respectively ( n = 6 per group). j , k Representative western blots of insulin-responsive Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) levels in the eWAT and skeletal muscle ( j ) and Akt and FOXO1 phosphorylation in the liver ( k ) of HFD-fed Redd1 KKAA mice and WT littermates ( n = 3). l Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 KKAA mice and WT littermates compared with NC-fed mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Weight gain in WT and Redd1 KKAA mice after being fed HFD for 16 weeks ( n = 10 per group). b eWAT and iWAT mass measurements in HFD-fed Redd1 KKAA mice and their WT littermates ( n = 10 per group). c Expression levels of Pparg and Cebpa in the eWAT of Redd1 KKAA mice and WT littermates fed HFD for 10 weeks ( n = 8 per group). d Representative images showing perilipin (green) and F4/80 (purple) staining in the eWAT of HFD-fed Redd1 KKAA mice and WT littermates ( n = 5 per group). Scale bar, 100 μm. e NF-κB activity in the eWAT from HFD-fed Redd1 KKAA mice and WT littermates ( n = 6 per group). f , g Plasma levels of inflammatory cytokines in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). h Fasting plasma levels of glucose and insulin in HFD-fed Redd1 KKAA mice and WT littermates ( n = 8 per group). i Assessment of GTT and ITT in HFD-fed Redd1 KKAA mice and WT littermates after fasting for 12 and 6 h, respectively ( n = 6 per group). j , k Representative western blots of insulin-responsive Akt phosphorylation and plasma membrane-associated GLUT4 (PM-GLUT4) levels in the eWAT and skeletal muscle ( j ) and Akt and FOXO1 phosphorylation in the liver ( k ) of HFD-fed Redd1 KKAA mice and WT littermates ( n = 3). l Relative expression levels of G6pc , Pck1 , and Fbp1 in the liver of HFD-fed Redd1 KKAA mice and WT littermates compared with NC-fed mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Expressing, Staining, Activity Assay, Clinical Proteomics, Western Blot, Phospho-proteomics, Membrane, Two Tailed Test

    a Representative images of H&E-stained liver tissues from HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and control mice, and quantification of hepatic steatosis from H&E-stained liver tissues ( n = 6 per group). Scale bars, 100 μm. b Expression levels of Acc , Fasn , and Scd-1 in the liver of HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and their control mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: REDD1 promotes obesity-induced metabolic dysfunction via atypical NF-κB activation

    doi: 10.1038/s41467-022-34110-1

    Figure Lengend Snippet: a Representative images of H&E-stained liver tissues from HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and control mice, and quantification of hepatic steatosis from H&E-stained liver tissues ( n = 6 per group). Scale bars, 100 μm. b Expression levels of Acc , Fasn , and Scd-1 in the liver of HFD-fed Redd1 −/− , Redd1 ΔAdipoq , Redd1 Δ LysM , Redd1 KKAA , and their control mice ( n = 6 per group). Bar graphs represent mean ± s.e.m. Statistical significance was calculated using an unpaired two-tailed t -test. Source data are provided as a Source Data file.

    Article Snippet: In addition, mouse adipose SVF cells and 3T3-L1 preadipocytes (5.0 × 10 5 cells/well, American Type Culture Collection) were plated and transfected with sh Redd1 (#sc-45807-SH, Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Staining, Control, Expressing, Two Tailed Test

    NFIL3 promotes the transcription of REDD1. (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 promotes the transcription of REDD1. (A) mRNA expression of REDD1 determined by RT-qPCR in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (B) Western blot analysis of REDD1 protein in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. *** p < 0.001, compared to healthy controls. (C) mRNA expression of REDD1 determined by RT-qPCR in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (D) Western blot analysis of REDD1 protein in MSU-treated neutrophils. ** p < 0.01, compared to PBS-treated neutrophils. (E) The enrichment of NFIL3 in the REDD1 promoter region detected by ChIP assay. ** p < 0.01, compared to IgG antibody. (F) A schematic diagram of NFIL3 binding sites in the REDD1 promoter region. (G) Binding of NFIL3 to the REDD1 promoter analyzed by dual-luciferase reporter assay. *** p < 0.001, compared to cells treated with pGL3-REDD1 + oe-NC. (H) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ### p < 0.001, compared to cells treated with Ad-NC. (I) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils following NFIL3 silencing or overexpression. ** p < 0.01 or *** p < 0.001, compared to cells treated with sh-NC. ## p < 0.01 or ### p < 0.001, compared to cells treated with Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Assay, Over Expression, Standard Deviation

    REDD1 inactivates the mTOR pathway to trigger neutrophil autophagy and inflammatory response. (A) REDD1-related genes analyzed by the GeneMANIA database. Each circle in the figure represents a gene, and the line between circles indicates interaction between two genes. (B) KEGG enrichment analysis of REDD1-related genes. The abscissa represents GeneRatio, the ordinate represents the entry identifier, the color represents enrichment p -value, and the size represents the number of enriched genes in the identifier. (C) Western blot analysis of p-S6 and p-4EBP in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. (D) Levels of IL-17, IL-23 and IL-1β in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. (E) Western blot analysis of p-S6 and p-4EBP in the MSU-treated neutrophils. (F) Levels of IL-17, IL-23 and IL-1β in the MSU-treated cell supernatant measured by ELISA. (G) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1. (H) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1. (I) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1 or combined with Rapamycin. (J) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1 or combined with Rapamycin. (K) Levels of IL-17, IL-23 and IL-1β in the supernatant of cells treated with sh-REDD1 or combined with Rapamycin measured by ELISA. In (C,D) , *** p < 0.001, compared to healthy controls. In (E,F) , ** p < 0.01, compared to cells treated with PBS; *** p < 0.001, compared to cells treated with PBS. In (G,H) , ** p < 0.01, compared to cells treated with sh-NC; *** p < 0.001, compared to cells treated with sh-NC. In (I–K) , ** p < 0.01, compared to cells treated with sh-NC + DMSO; ## p < 0.01, compared to cells treated with sh-REDD1 + DMSO; ### p < 0.001, compared to cells treated with sh-REDD1 + DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: REDD1 inactivates the mTOR pathway to trigger neutrophil autophagy and inflammatory response. (A) REDD1-related genes analyzed by the GeneMANIA database. Each circle in the figure represents a gene, and the line between circles indicates interaction between two genes. (B) KEGG enrichment analysis of REDD1-related genes. The abscissa represents GeneRatio, the ordinate represents the entry identifier, the color represents enrichment p -value, and the size represents the number of enriched genes in the identifier. (C) Western blot analysis of p-S6 and p-4EBP in the peripheral blood neutrophils of 28 gout patients and 20 healthy controls. (D) Levels of IL-17, IL-23 and IL-1β in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. (E) Western blot analysis of p-S6 and p-4EBP in the MSU-treated neutrophils. (F) Levels of IL-17, IL-23 and IL-1β in the MSU-treated cell supernatant measured by ELISA. (G) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1. (H) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1. (I) mRNA expression of REDD1 determined by RT-qPCR in neutrophils treated with sh-REDD1 or combined with Rapamycin. (J) Western blot analysis of p-S6, p-4EBP and LC3 II in neutrophils treated with sh-REDD1 or combined with Rapamycin. (K) Levels of IL-17, IL-23 and IL-1β in the supernatant of cells treated with sh-REDD1 or combined with Rapamycin measured by ELISA. In (C,D) , *** p < 0.001, compared to healthy controls. In (E,F) , ** p < 0.01, compared to cells treated with PBS; *** p < 0.001, compared to cells treated with PBS. In (G,H) , ** p < 0.01, compared to cells treated with sh-NC; *** p < 0.001, compared to cells treated with sh-NC. In (I–K) , ** p < 0.01, compared to cells treated with sh-NC + DMSO; ## p < 0.01, compared to cells treated with sh-REDD1 + DMSO; ### p < 0.001, compared to cells treated with sh-REDD1 + DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Standard Deviation

    REDD1 induces the formation of NETs by promoting neutrophil autophagy. (A) Levels of NETs in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. *** p < 0.001, compared to healthy controls. (B) Immunofluorescence staining analysis of the formation of NETs in MSU-treated neutrophils, scale bar = 50 μm. Neutrophil elastase (NE) and DAPI (DNA) were used to stain the cells. The white arrow represents the formed NETs. *** p < 0.001, compared to PBS-treated neutrophils. (C) Western blot analysis of LC3 II protein in MSU-induced neutrophils treated with Baf A1. ** p < 0.001, compared to MSU-treated neutrophils. (D) Immunofluorescence staining analysis of the formation of NETs in MSU-induced neutrophils treated with Baf A1. ** p < 0.01, compared to MSU-treated neutrophils. (E) Immunofluorescence staining analysis of the formation of NETs in neutrophils treated with sh-REDD1 or combined with Rapamycin, scale bar = 50 μm. ** p < 0.01, compared to neutrophils treated with sh-NC and DMSO. ## p < 0.01, compared to neutrophils treated with sh-REDD1 and DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: REDD1 induces the formation of NETs by promoting neutrophil autophagy. (A) Levels of NETs in the peripheral blood of 28 gout patients and 20 healthy controls measured by ELISA. *** p < 0.001, compared to healthy controls. (B) Immunofluorescence staining analysis of the formation of NETs in MSU-treated neutrophils, scale bar = 50 μm. Neutrophil elastase (NE) and DAPI (DNA) were used to stain the cells. The white arrow represents the formed NETs. *** p < 0.001, compared to PBS-treated neutrophils. (C) Western blot analysis of LC3 II protein in MSU-induced neutrophils treated with Baf A1. ** p < 0.001, compared to MSU-treated neutrophils. (D) Immunofluorescence staining analysis of the formation of NETs in MSU-induced neutrophils treated with Baf A1. ** p < 0.01, compared to MSU-treated neutrophils. (E) Immunofluorescence staining analysis of the formation of NETs in neutrophils treated with sh-REDD1 or combined with Rapamycin, scale bar = 50 μm. ** p < 0.01, compared to neutrophils treated with sh-NC and DMSO. ## p < 0.01, compared to neutrophils treated with sh-REDD1 and DMSO. Data are shown as mean ± standard deviation of three technical replicates. Data between two groups were compared using unpaired t -test while those among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Western Blot, Standard Deviation

    NFIL3 silencing contributes to suppressed neutrophil autophagy and NET formation, which is related to downregulated expression of REDD1. (A) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (B) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (C) Western blot analysis of p-S6 and p-4EBP1 in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (D) Western blot analysis of LC3 II protein in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (E) NET formation in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1, scale bar = 50 μm. *** p < 0.001, compared to neutrophils treated with sh-NC + Ad-NC. ## p < 0.01, compared to neutrophils treated with sh-NFIL3 + Ad-NC. ### p < 0.001, compared to neutrophils treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 silencing contributes to suppressed neutrophil autophagy and NET formation, which is related to downregulated expression of REDD1. (A) mRNA expression of NFIL3 and REDD1 determined by RT-qPCR in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (B) Western blot analysis of NFIL3 and REDD1 proteins in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (C) Western blot analysis of p-S6 and p-4EBP1 in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (D) Western blot analysis of LC3 II protein in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1. (E) NET formation in neutrophils treated with sh-NFIL3 or combined with Ad-REDD1, scale bar = 50 μm. *** p < 0.001, compared to neutrophils treated with sh-NC + Ad-NC. ## p < 0.01, compared to neutrophils treated with sh-NFIL3 + Ad-NC. ### p < 0.001, compared to neutrophils treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

    NFIL3 silencing impairs neutrophil autophagy, reduces NET formation and inflammatory injury in mice with acute gouty arthritis via the REDD1/mTOR axis. PBS-treated mice served as the control and MSU mice were treated with sh-NFIL3 or combined with Ad-REDD1. (A) Representative microscopic views of ankle joint swelling of mice (the left), and the corresponding quantitative analysis (the right). (B) The representative images of pathological changes in the joint tissues of mice following HE staining, scale bar = 25 μm. (C) mRNA expression of NFIL3 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (D) mRNA expression of REDD1 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (E) Western blot analysis of NFIL3 expression in the neutrophils of the joint tissues of mice. (F) Quantitative analysis of REDD1 protein expression. (G) Quantitative analysis of S6 and 4EBP phosphorylation levels. (H) Quantitative analysis of LC3 II protein expression. (I) MPO activity in the joint tissue of mice. (J) IL-17 levels measured by ELISA in the serum of mice. (K) IL-23 levels measured by ELISA in the serum of mice. (L) IL-1β levels measured by ELISA in the serum of mice. (M) Quantitative analysis of NET formation in the joint tissue of mice. *** p < 0.001, compared to PBS-treated mice. ## p < 0.01, compared to MSU mice treated with sh-NC + Ad-NC. ### p < 0.001, compared to MSU mice treated with sh-NC + Ad-NC. &, && or &&& p < 0.05, compared to MSU mice treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA. n = 10 for mice in each group.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: NFIL3 silencing impairs neutrophil autophagy, reduces NET formation and inflammatory injury in mice with acute gouty arthritis via the REDD1/mTOR axis. PBS-treated mice served as the control and MSU mice were treated with sh-NFIL3 or combined with Ad-REDD1. (A) Representative microscopic views of ankle joint swelling of mice (the left), and the corresponding quantitative analysis (the right). (B) The representative images of pathological changes in the joint tissues of mice following HE staining, scale bar = 25 μm. (C) mRNA expression of NFIL3 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (D) mRNA expression of REDD1 determined by RT-qPCR in the neutrophils of the joint tissue of mice. (E) Western blot analysis of NFIL3 expression in the neutrophils of the joint tissues of mice. (F) Quantitative analysis of REDD1 protein expression. (G) Quantitative analysis of S6 and 4EBP phosphorylation levels. (H) Quantitative analysis of LC3 II protein expression. (I) MPO activity in the joint tissue of mice. (J) IL-17 levels measured by ELISA in the serum of mice. (K) IL-23 levels measured by ELISA in the serum of mice. (L) IL-1β levels measured by ELISA in the serum of mice. (M) Quantitative analysis of NET formation in the joint tissue of mice. *** p < 0.001, compared to PBS-treated mice. ## p < 0.01, compared to MSU mice treated with sh-NC + Ad-NC. ### p < 0.001, compared to MSU mice treated with sh-NC + Ad-NC. &, && or &&& p < 0.05, compared to MSU mice treated with sh-NFIL3 + Ad-NC. Data are shown as mean ± standard deviation of three technical replicates. Data among multiple groups were assessed by one-way ANOVA. n = 10 for mice in each group.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: Control, Staining, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Schematic diagram of the mechanism by which NFIL3 affects gout. NFIL3 promotes the transcription of REDD1 and inhibits the mTOR pathway, thus activating the autophagy of neutrophils and inducing the formation of NETs, eventually aggravating the inflammatory response of gout.

    Journal: Frontiers in Medicine

    Article Title: NFIL3 Facilitates Neutrophil Autophagy, Neutrophil Extracellular Trap Formation and Inflammation During Gout via REDD1-Dependent mTOR Inactivation

    doi: 10.3389/fmed.2021.692781

    Figure Lengend Snippet: Schematic diagram of the mechanism by which NFIL3 affects gout. NFIL3 promotes the transcription of REDD1 and inhibits the mTOR pathway, thus activating the autophagy of neutrophils and inducing the formation of NETs, eventually aggravating the inflammatory response of gout.

    Article Snippet: Next, the cells were treated with sh-NC, sh-NFIL3, Ad-NC, Ad-NFIL3, sh-REDD1, sh-NC + dimethyl sulphoxide (DMSO), sh-REDD1 + DMSO, sh-REDD1 + Rapamycin (mTOR inhibitor; MedChemExpress Company, USA; 100 nM), sh-NC + Ad-NC, sh-NFIL3 + Ad-NC and sh-NFIL3 + Ad-REDD1.

    Techniques: