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recombinant mouse il10  (Sino Biological)


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    Sino Biological recombinant mouse il10
    a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of <t>Il10</t> in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.
    Recombinant Mouse Il10, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mouse il10/product/Sino Biological
    Average 94 stars, based on 7 article reviews
    recombinant mouse il10 - by Bioz Stars, 2026-03
    94/100 stars

    Images

    1) Product Images from "Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression"

    Article Title: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression

    Journal: Nature Communications

    doi: 10.1038/s41467-025-61371-3

    a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of Il10 in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.
    Figure Legend Snippet: a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of Il10 in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.

    Techniques Used: Quantitative RT-PCR, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Western Blot, Isolation, Membrane, Luciferase, Activity Assay, Over Expression

    a qRT-PCR analysis of Il10 in iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 8. b Western blot analysis with densitometric quantification of IL10 in BAT and iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). c IL10 ELISA assays in culture medium collected from adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 5). d , e qRT-PCR analysis of IL10 target genes in differentiated brown ( d ) and beige ( e ) adipocytes treated with dbcAMP while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 4). f IL10 ELISA assays in culture medium collected from AAV-transfected adipocytes isolated from wild-type mice ( n = 4). g – j qRT-PCR analysis of IL10 target genes ( g and h , n = 3) and OCR ( i and j ) in differentiated brown and beige adipocytes treated with dbcAMP along with either an IL10 neutralizing antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice. i , IgG ( n = 3), anti-IL10 ( n = 4). j , IgG ( n = 4), anti-IL10 ( n = 5). k , l qRT-PCR analysis of IL10 target genes in differentiated brown ( k ) and beige ( l ) adipocytes treated with dbcAMP along with either an IL10Rα blocking antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided Mann–Whitney test (BAT in c ) or two-sided unpaired t test ( a , b , iWAT in c , d – l ). Ads, Adipocytes. Source data are provided as a Source Data file.
    Figure Legend Snippet: a qRT-PCR analysis of Il10 in iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 8. b Western blot analysis with densitometric quantification of IL10 in BAT and iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). c IL10 ELISA assays in culture medium collected from adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 5). d , e qRT-PCR analysis of IL10 target genes in differentiated brown ( d ) and beige ( e ) adipocytes treated with dbcAMP while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 4). f IL10 ELISA assays in culture medium collected from AAV-transfected adipocytes isolated from wild-type mice ( n = 4). g – j qRT-PCR analysis of IL10 target genes ( g and h , n = 3) and OCR ( i and j ) in differentiated brown and beige adipocytes treated with dbcAMP along with either an IL10 neutralizing antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice. i , IgG ( n = 3), anti-IL10 ( n = 4). j , IgG ( n = 4), anti-IL10 ( n = 5). k , l qRT-PCR analysis of IL10 target genes in differentiated brown ( k ) and beige ( l ) adipocytes treated with dbcAMP along with either an IL10Rα blocking antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided Mann–Whitney test (BAT in c ) or two-sided unpaired t test ( a , b , iWAT in c , d – l ). Ads, Adipocytes. Source data are provided as a Source Data file.

    Techniques Used: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Isolation, Cell Culture, Transfection, Control, Blocking Assay, MANN-WHITNEY

    a , b Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( a ) or iWAT ( b ) injections of Il10ra siRNA (si- Il10ra ) or control siRNA (si- Ctrl ), maintained at 4 °C for 7 days. c – g Analysis in BAT after siRNA injections: qRT-PCR analysis of Il10ra ( c , n = 5), Western blot analysis with densitometric quantification of PGC1α, IL10Rα, and UCP1 ( d , n = 3), qRT-PCR analysis of IL10 target genes ( e , n = 5), HE staining with lipid droplet size quantification ( f , n = 5), and rectal temperature ( g , n = 7 for si- Ctrl , n = 9 for si- Il10ra ). h - l Analysis in iWAT after siRNA injections: qRT-PCR analysis of Il10ra ( h , n = 5), Western blot with densitometric quantification analyses of PGC1α, IL10Rα, and UCP1 ( i , n = 3), qRT-PCR analyses of IL10 target genes ( j , n = 5), HE staining with lipid droplet size quantification ( k , n = 5), and rectal temperature ( l , n = 5 for si- Ctrl , n = 7 for si- Il10ra ). m, n Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( m ) or iWAT ( n ) injections of AAV-sh- Ctrl or AAV-sh- Il10 , housed for 3 weeks before 7 days at 4 °C. o - s Analysis in BAT AAV-injected mice: qRT-PCR analysis of Il10 in BAT, liver, and muscle ( o , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( p , n = 3), qRT-PCR analysis of IL10 target genes ( q , n = 6), HE staining with lipid droplet size quantification ( r , n = 5), rectal temperature ( s , n = 10). t –x Analysis in iWAT AAV-injected mice: qRT-PCR analysis of Il10 in iWAT, liver, and muscle ( t , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( u , n = 3), qRT-PCR analysis of IL10 target genes in iWAT ( v , n = 6 for AAV-sh- Ctrl , n = 7 for AAV-sh- Il10 ), HE staining with lipid droplet size quantification ( w , n = 5), and rectal temperature ( x , n = 10). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided unpaired t test. Source data are provided as a Source Data file. Panels a and m were created in BioRender . Panels b and n were created in BioRender .
    Figure Legend Snippet: a , b Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( a ) or iWAT ( b ) injections of Il10ra siRNA (si- Il10ra ) or control siRNA (si- Ctrl ), maintained at 4 °C for 7 days. c – g Analysis in BAT after siRNA injections: qRT-PCR analysis of Il10ra ( c , n = 5), Western blot analysis with densitometric quantification of PGC1α, IL10Rα, and UCP1 ( d , n = 3), qRT-PCR analysis of IL10 target genes ( e , n = 5), HE staining with lipid droplet size quantification ( f , n = 5), and rectal temperature ( g , n = 7 for si- Ctrl , n = 9 for si- Il10ra ). h - l Analysis in iWAT after siRNA injections: qRT-PCR analysis of Il10ra ( h , n = 5), Western blot with densitometric quantification analyses of PGC1α, IL10Rα, and UCP1 ( i , n = 3), qRT-PCR analyses of IL10 target genes ( j , n = 5), HE staining with lipid droplet size quantification ( k , n = 5), and rectal temperature ( l , n = 5 for si- Ctrl , n = 7 for si- Il10ra ). m, n Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( m ) or iWAT ( n ) injections of AAV-sh- Ctrl or AAV-sh- Il10 , housed for 3 weeks before 7 days at 4 °C. o - s Analysis in BAT AAV-injected mice: qRT-PCR analysis of Il10 in BAT, liver, and muscle ( o , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( p , n = 3), qRT-PCR analysis of IL10 target genes ( q , n = 6), HE staining with lipid droplet size quantification ( r , n = 5), rectal temperature ( s , n = 10). t –x Analysis in iWAT AAV-injected mice: qRT-PCR analysis of Il10 in iWAT, liver, and muscle ( t , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( u , n = 3), qRT-PCR analysis of IL10 target genes in iWAT ( v , n = 6 for AAV-sh- Ctrl , n = 7 for AAV-sh- Il10 ), HE staining with lipid droplet size quantification ( w , n = 5), and rectal temperature ( x , n = 10). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided unpaired t test. Source data are provided as a Source Data file. Panels a and m were created in BioRender . Panels b and n were created in BioRender .

    Techniques Used: In Situ, Control, Quantitative RT-PCR, Western Blot, Staining, Injection

    CLDN5 is highly enriched in the UCP1 low-expressing adipocytes subpopulation. CLDN5 forms a complex with ZO1 and YBX3 in normal adipocytes under physiological conditions. Mice lacking CLDN5 in adipocytes have reduced heat production, reduced energy consumption, and an obesity-prone phenotype. CLDN5 deletion causes YBX3 to enter the nucleus, promotes the expression and secretion of IL10, and then affects the expression of thermogenic genes in neighboring thermogenic cells through IL10-IL10Ra signaling. The graphic was created in BioRender .
    Figure Legend Snippet: CLDN5 is highly enriched in the UCP1 low-expressing adipocytes subpopulation. CLDN5 forms a complex with ZO1 and YBX3 in normal adipocytes under physiological conditions. Mice lacking CLDN5 in adipocytes have reduced heat production, reduced energy consumption, and an obesity-prone phenotype. CLDN5 deletion causes YBX3 to enter the nucleus, promotes the expression and secretion of IL10, and then affects the expression of thermogenic genes in neighboring thermogenic cells through IL10-IL10Ra signaling. The graphic was created in BioRender .

    Techniques Used: Expressing



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    a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of <t>Il10</t> in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.
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    Thermo Fisher mouse il10 recombinant protein, ebiosciencetm
    Figure 3. GCN2 levels regulate macrophage polarization during skin wound healing (A) Flow cytometry analysis of F4/80+CD11b+ skin wound macrophages (gated on CD45+ cells) from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (B) Flow cytometry analysis of CD86 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (C) Flow cytometry analysis of CD206 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (D) CD206, <t>Il10,</t> Tgfb1, and Vegfa expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (E) Cx3cr1 and Il6 expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (F and G) Rose diagrams showing all highly expressed GCN2KO genes and all highly expressed WT genes at 3 (F) and 5 dpi (G). (H–J) Heatmap showing differentially expressed M1 (H), M2a (I), and M2c (J) marker genes in WT and GCN2KO mice. Macrophages were obtained from sorted skin wound macrophages (A–G) or peritoneal macrophages (H–J). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S3.
    Mouse Il10 Recombinant Protein, Ebiosciencetm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems cf recombinant il10 r d systems
    Figure 3. GCN2 levels regulate macrophage polarization during skin wound healing (A) Flow cytometry analysis of F4/80+CD11b+ skin wound macrophages (gated on CD45+ cells) from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (B) Flow cytometry analysis of CD86 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (C) Flow cytometry analysis of CD206 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (D) CD206, <t>Il10,</t> Tgfb1, and Vegfa expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (E) Cx3cr1 and Il6 expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (F and G) Rose diagrams showing all highly expressed GCN2KO genes and all highly expressed WT genes at 3 (F) and 5 dpi (G). (H–J) Heatmap showing differentially expressed M1 (H), M2a (I), and M2c (J) marker genes in WT and GCN2KO mice. Macrophages were obtained from sorted skin wound macrophages (A–G) or peritoneal macrophages (H–J). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S3.
    Cf Recombinant Il10 R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cf recombinant il10 r d systems/product/R&D Systems
    Average 96 stars, based on 1 article reviews
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    a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of Il10 in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression

    doi: 10.1038/s41467-025-61371-3

    Figure Lengend Snippet: a qRT-PCR analysis of Zo1 in GFP + and GFP - adipocytes from BAT/iWAT of CLDN5-GFP mice ( n = 3). b Co-IP showing that endogenous CLDN5 interacts with endogenous ZO1/YBX3 in BAT/iWAT. IP/Blot antibodies as indicated. *Lanes show 10% of the input amount of other lanes. c Immunofluorescence staining of YBX3 in BAT/iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. Scale bar, BAT (20 μm) and iWAT (50 μm). d Western blot analysis of the subcellular localization of YBX3 in adipocytes isolated from BAT/iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice. ATP1A, GAPDH, and Histone H3 serving as membrane fraction (M), cytosolic fraction (C), and nuclear fraction (N) controls. e qRT-PCR analysis of Il10 in BAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 6. f , g qRT-PCR analysis ( f , in BAT, n = 7 for RT or 7 d, n = 8 for 6 h; in iWAT, n = 8.) and Western blot analysis with densitometric quantification ( g , n = 3) of IL10 in BAT and iWAT of wild-type mice housed at RT, 4 °C for 6 h or 7 days (7 d). h – k qRT-PCR ( h and j , n = 3) and Western blot ( i and k , n = 3) analysis of IL10 in dbcAMP-stimulated brown/beige adipocytes from wild-type mice harvested at the indicated time points. l - m Association of YBX3 with Il10 mRNA as measured by RIP and qRT-PCR analysis from wild-type mice using an YBX3/IgG antibody ( l ) or from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice using an anti-YBX3 antibody ( m ), n = 3 biological replicates. n – p Luciferase assays in HEK293 cells. Il10 3’UTR/promoter activity with YBX3 overexpression ( n and p ); 3’UTR deletion mutants with YBX3 overexpression ( o ); n = 4 independent experiments. Statistical analyses were performed using two-sided unpaired t test ( a , e , l and m ), two-sided one-way ANOVA with Dunnett’s post-hoc test ( f – h , j ) or Tukey’s post-hoc test ( n – p ). For b – d , each experiment was repeated three times with consistent results. Data are expressed as the mean ± SEM. Ads, Adipocytes. Source data are provided as a Source Data file.

    Article Snippet: 10-week-old wild-type mice were randomly allocated into four experimental groups and received bilateral, multi-point injections of either PBS (vehicle control) or recombinant mouse IL10 (SinoBiological, 50245-MNAE) at doses of 50, 250, or 500 ng in 40 μL PBS into the interscapular BAT or bilateral iWAT depots.

    Techniques: Quantitative RT-PCR, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Western Blot, Isolation, Membrane, Luciferase, Activity Assay, Over Expression

    a qRT-PCR analysis of Il10 in iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 8. b Western blot analysis with densitometric quantification of IL10 in BAT and iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). c IL10 ELISA assays in culture medium collected from adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 5). d , e qRT-PCR analysis of IL10 target genes in differentiated brown ( d ) and beige ( e ) adipocytes treated with dbcAMP while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 4). f IL10 ELISA assays in culture medium collected from AAV-transfected adipocytes isolated from wild-type mice ( n = 4). g – j qRT-PCR analysis of IL10 target genes ( g and h , n = 3) and OCR ( i and j ) in differentiated brown and beige adipocytes treated with dbcAMP along with either an IL10 neutralizing antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice. i , IgG ( n = 3), anti-IL10 ( n = 4). j , IgG ( n = 4), anti-IL10 ( n = 5). k , l qRT-PCR analysis of IL10 target genes in differentiated brown ( k ) and beige ( l ) adipocytes treated with dbcAMP along with either an IL10Rα blocking antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided Mann–Whitney test (BAT in c ) or two-sided unpaired t test ( a , b , iWAT in c , d – l ). Ads, Adipocytes. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression

    doi: 10.1038/s41467-025-61371-3

    Figure Lengend Snippet: a qRT-PCR analysis of Il10 in iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice, n = 8. b Western blot analysis with densitometric quantification of IL10 in BAT and iWAT from Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). c IL10 ELISA assays in culture medium collected from adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 5). d , e qRT-PCR analysis of IL10 target genes in differentiated brown ( d ) and beige ( e ) adipocytes treated with dbcAMP while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox and Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 4). f IL10 ELISA assays in culture medium collected from AAV-transfected adipocytes isolated from wild-type mice ( n = 4). g – j qRT-PCR analysis of IL10 target genes ( g and h , n = 3) and OCR ( i and j ) in differentiated brown and beige adipocytes treated with dbcAMP along with either an IL10 neutralizing antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice. i , IgG ( n = 3), anti-IL10 ( n = 4). j , IgG ( n = 4), anti-IL10 ( n = 5). k , l qRT-PCR analysis of IL10 target genes in differentiated brown ( k ) and beige ( l ) adipocytes treated with dbcAMP along with either an IL10Rα blocking antibody or control IgG while co-cultured with CD36 + primary adipocytes isolated from BAT and iWAT of Cldn5 flox/flox ; Fabp4 -Cre mice ( n = 3). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided Mann–Whitney test (BAT in c ) or two-sided unpaired t test ( a , b , iWAT in c , d – l ). Ads, Adipocytes. Source data are provided as a Source Data file.

    Article Snippet: 10-week-old wild-type mice were randomly allocated into four experimental groups and received bilateral, multi-point injections of either PBS (vehicle control) or recombinant mouse IL10 (SinoBiological, 50245-MNAE) at doses of 50, 250, or 500 ng in 40 μL PBS into the interscapular BAT or bilateral iWAT depots.

    Techniques: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Isolation, Cell Culture, Transfection, Control, Blocking Assay, MANN-WHITNEY

    a , b Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( a ) or iWAT ( b ) injections of Il10ra siRNA (si- Il10ra ) or control siRNA (si- Ctrl ), maintained at 4 °C for 7 days. c – g Analysis in BAT after siRNA injections: qRT-PCR analysis of Il10ra ( c , n = 5), Western blot analysis with densitometric quantification of PGC1α, IL10Rα, and UCP1 ( d , n = 3), qRT-PCR analysis of IL10 target genes ( e , n = 5), HE staining with lipid droplet size quantification ( f , n = 5), and rectal temperature ( g , n = 7 for si- Ctrl , n = 9 for si- Il10ra ). h - l Analysis in iWAT after siRNA injections: qRT-PCR analysis of Il10ra ( h , n = 5), Western blot with densitometric quantification analyses of PGC1α, IL10Rα, and UCP1 ( i , n = 3), qRT-PCR analyses of IL10 target genes ( j , n = 5), HE staining with lipid droplet size quantification ( k , n = 5), and rectal temperature ( l , n = 5 for si- Ctrl , n = 7 for si- Il10ra ). m, n Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( m ) or iWAT ( n ) injections of AAV-sh- Ctrl or AAV-sh- Il10 , housed for 3 weeks before 7 days at 4 °C. o - s Analysis in BAT AAV-injected mice: qRT-PCR analysis of Il10 in BAT, liver, and muscle ( o , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( p , n = 3), qRT-PCR analysis of IL10 target genes ( q , n = 6), HE staining with lipid droplet size quantification ( r , n = 5), rectal temperature ( s , n = 10). t –x Analysis in iWAT AAV-injected mice: qRT-PCR analysis of Il10 in iWAT, liver, and muscle ( t , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( u , n = 3), qRT-PCR analysis of IL10 target genes in iWAT ( v , n = 6 for AAV-sh- Ctrl , n = 7 for AAV-sh- Il10 ), HE staining with lipid droplet size quantification ( w , n = 5), and rectal temperature ( x , n = 10). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided unpaired t test. Source data are provided as a Source Data file. Panels a and m were created in BioRender . Panels b and n were created in BioRender .

    Journal: Nature Communications

    Article Title: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression

    doi: 10.1038/s41467-025-61371-3

    Figure Lengend Snippet: a , b Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( a ) or iWAT ( b ) injections of Il10ra siRNA (si- Il10ra ) or control siRNA (si- Ctrl ), maintained at 4 °C for 7 days. c – g Analysis in BAT after siRNA injections: qRT-PCR analysis of Il10ra ( c , n = 5), Western blot analysis with densitometric quantification of PGC1α, IL10Rα, and UCP1 ( d , n = 3), qRT-PCR analysis of IL10 target genes ( e , n = 5), HE staining with lipid droplet size quantification ( f , n = 5), and rectal temperature ( g , n = 7 for si- Ctrl , n = 9 for si- Il10ra ). h - l Analysis in iWAT after siRNA injections: qRT-PCR analysis of Il10ra ( h , n = 5), Western blot with densitometric quantification analyses of PGC1α, IL10Rα, and UCP1 ( i , n = 3), qRT-PCR analyses of IL10 target genes ( j , n = 5), HE staining with lipid droplet size quantification ( k , n = 5), and rectal temperature ( l , n = 5 for si- Ctrl , n = 7 for si- Il10ra ). m, n Schematic of experimental design in Cldn5 flox/flox ; Fabp4 -Cre mice, with in situ BAT ( m ) or iWAT ( n ) injections of AAV-sh- Ctrl or AAV-sh- Il10 , housed for 3 weeks before 7 days at 4 °C. o - s Analysis in BAT AAV-injected mice: qRT-PCR analysis of Il10 in BAT, liver, and muscle ( o , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( p , n = 3), qRT-PCR analysis of IL10 target genes ( q , n = 6), HE staining with lipid droplet size quantification ( r , n = 5), rectal temperature ( s , n = 10). t –x Analysis in iWAT AAV-injected mice: qRT-PCR analysis of Il10 in iWAT, liver, and muscle ( t , n = 6), Western blot analysis with densitometric quantification of IL10, PGC1α, and UCP1 ( u , n = 3), qRT-PCR analysis of IL10 target genes in iWAT ( v , n = 6 for AAV-sh- Ctrl , n = 7 for AAV-sh- Il10 ), HE staining with lipid droplet size quantification ( w , n = 5), and rectal temperature ( x , n = 10). Data are expressed as the mean ± SEM. Statistical analyses were performed using two-sided unpaired t test. Source data are provided as a Source Data file. Panels a and m were created in BioRender . Panels b and n were created in BioRender .

    Article Snippet: 10-week-old wild-type mice were randomly allocated into four experimental groups and received bilateral, multi-point injections of either PBS (vehicle control) or recombinant mouse IL10 (SinoBiological, 50245-MNAE) at doses of 50, 250, or 500 ng in 40 μL PBS into the interscapular BAT or bilateral iWAT depots.

    Techniques: In Situ, Control, Quantitative RT-PCR, Western Blot, Staining, Injection

    CLDN5 is highly enriched in the UCP1 low-expressing adipocytes subpopulation. CLDN5 forms a complex with ZO1 and YBX3 in normal adipocytes under physiological conditions. Mice lacking CLDN5 in adipocytes have reduced heat production, reduced energy consumption, and an obesity-prone phenotype. CLDN5 deletion causes YBX3 to enter the nucleus, promotes the expression and secretion of IL10, and then affects the expression of thermogenic genes in neighboring thermogenic cells through IL10-IL10Ra signaling. The graphic was created in BioRender .

    Journal: Nature Communications

    Article Title: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression

    doi: 10.1038/s41467-025-61371-3

    Figure Lengend Snippet: CLDN5 is highly enriched in the UCP1 low-expressing adipocytes subpopulation. CLDN5 forms a complex with ZO1 and YBX3 in normal adipocytes under physiological conditions. Mice lacking CLDN5 in adipocytes have reduced heat production, reduced energy consumption, and an obesity-prone phenotype. CLDN5 deletion causes YBX3 to enter the nucleus, promotes the expression and secretion of IL10, and then affects the expression of thermogenic genes in neighboring thermogenic cells through IL10-IL10Ra signaling. The graphic was created in BioRender .

    Article Snippet: 10-week-old wild-type mice were randomly allocated into four experimental groups and received bilateral, multi-point injections of either PBS (vehicle control) or recombinant mouse IL10 (SinoBiological, 50245-MNAE) at doses of 50, 250, or 500 ng in 40 μL PBS into the interscapular BAT or bilateral iWAT depots.

    Techniques: Expressing

    Figure 3. GCN2 levels regulate macrophage polarization during skin wound healing (A) Flow cytometry analysis of F4/80+CD11b+ skin wound macrophages (gated on CD45+ cells) from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (B) Flow cytometry analysis of CD86 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (C) Flow cytometry analysis of CD206 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (D) CD206, Il10, Tgfb1, and Vegfa expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (E) Cx3cr1 and Il6 expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (F and G) Rose diagrams showing all highly expressed GCN2KO genes and all highly expressed WT genes at 3 (F) and 5 dpi (G). (H–J) Heatmap showing differentially expressed M1 (H), M2a (I), and M2c (J) marker genes in WT and GCN2KO mice. Macrophages were obtained from sorted skin wound macrophages (A–G) or peritoneal macrophages (H–J). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S3.

    Journal: Cell reports

    Article Title: Downregulation of nutrition sensor GCN2 in macrophages contributes to poor wound healing in diabetes.

    doi: 10.1016/j.celrep.2023.113658

    Figure Lengend Snippet: Figure 3. GCN2 levels regulate macrophage polarization during skin wound healing (A) Flow cytometry analysis of F4/80+CD11b+ skin wound macrophages (gated on CD45+ cells) from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (B) Flow cytometry analysis of CD86 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (C) Flow cytometry analysis of CD206 expression in skin wound macrophages from WT and GCN2KO mice at 3 or 5 dpi (n = 3). (D) CD206, Il10, Tgfb1, and Vegfa expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (E) Cx3cr1 and Il6 expression in 3 or 5 dpi skin macrophages from WT and GCN2KO mice analyzed by real-time PCR (n = 3). (F and G) Rose diagrams showing all highly expressed GCN2KO genes and all highly expressed WT genes at 3 (F) and 5 dpi (G). (H–J) Heatmap showing differentially expressed M1 (H), M2a (I), and M2c (J) marker genes in WT and GCN2KO mice. Macrophages were obtained from sorted skin wound macrophages (A–G) or peritoneal macrophages (H–J). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S3.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies FITC anti-mouse CD206 BioLegend Cat# 141703; RRID: AB_10900988 PE anti-mouse F4/80 BioLegend Cat# 123110; RRID: AB_893486 APC anti-mouse F4/80 Thermo Fisher Cat# 17-4801-82; RRID: AB_2784648 PE/Cy5 anti-mouse CD11b Thermo Fisher Cat# 15-0112-83; RRID: AB_468714 Bv510 anti-mouse CD11b BioLegend Cat# 101263; RRID: AB_2629529 PE anti-mouse CD124 BioLegend Cat# 144804; RRID: AB_2561730 FITC anti-mouse F4/80 Thermo Fisher Cat# 11-4801-82; RRID: AB_2637191 PE anti-mouse TNFa Thermo Fisher Cat# 12-7321-82; RRID: AB_466199 FITC anti-mouse CD86 eBioscience Cat# 105006; RRID: AB_313149 PE/Cy5 anti-mouse CD45 Thermo Fisher Cat# 15-0459-42; RRID: AB_10717815 APC anti-mouse CD45 BioLegend Cat# 103112; RRID: AB_312977 PE anti-mouse Ly6G BioLegend Cat# 127607; RRID: AB_1186104 PE anti-mouse CD115 BioLegend Cat# 135506; RRID: AB_1937253 PE/Cy7 anti-mouse CD45.1 BioLegend Cat# 110730; RRID: AB_1134168 Fixable Viability Dye eFluorTM 780 Thermo Fisher Cat# 65-0865-14; RRID: —— PERCP/CY5.5 anti-mouse CD45.2 BioLegend Cat# 109827; RRID: AB_893352 Brilliant Violet 510 anti-mouse CD11b BioLegend Cat# 101245; RRID: AB_2561390 Brilliant Violet 605 anti-mouse Ly6C BioLegend Cat# 128036; RRID: AB_2562353 b-actin Sigma Cat# A5441; RRID: AB_476744 Arginase1 Santa Cruz Cat# sc-20150; RRID: AB_2058955 GCN2 Cell Signaling Technology Cat# 3302s; RRID: AB_10694800 p-IkBa Cell Signaling Technology Cat# 92456s; RRID: AB_2267145 IkBa Abcam Cat# ab32518; RRID: AB_2801653 p-IKKa/b Cell Signaling Technology Cat# 14938; RRID: AB_2798653 IKKa Cell Signaling Technology Cat# 61294s; RRID: AB_2799606 P65 Cell Signaling Technology Cat# 6596s; RRID: AB_2128043 p-P65 Cell Signaling Technology Cat# 3033T; RRID: AB_331284 Hif1a Cell Signaling Technology Cat# 36169; RRID: AB_2799095 p-eIF2a Cell Signaling Technology Cat# 9721s; RRID: AB_330951 eIF2a Cell Signaling Technology Cat# 9722s; RRID: AB_2230924 p-GCN2 Abcam Cat# ab75836; RRID: AB_1310260 Chemicals, peptides, and recombinant proteins Recombinant Murine M-CSF Pepro Tech Cat# 315-02 Thioglycollate BD BBL Cat# 211716 Recombinant murine IL4 R&D Systems Cat# 404-ML Trizol Thermo Fisher Cat# 15596018 Recombinant murine IL10 R&D Systems Cat# 417-ML Recombinant murine IFN-g R&D Systems Cat# 485-ML Lipopolysaccharide Sigma Cat# L2630 SYBR Green Mixture TaKaRa Cat# RR420A AMV Reverse transcriptase TaKaRa Cat# 2621 Oligo dT18 Thermo Fisher Cat# 18418020 Recombinant RNase Inhibitor TaKaRa Cat# 2313A (Continued on next page) 16 Cell Reports 43, 113658, January 23, 2024

    Techniques: Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Marker

    Figure 6. GCN2 is involved in T1D mouse delayed skin wound healing (A) Wound-healing pictures from control, T1D WT, and GCN2KO mice. (B) Wound-healing rates in the control group, WT T1D group, and GCN2KO group (n = 5). (C) Skin tissue H&E staining from the control group, WT T1D group, and GCN2KO group at 7 dpi (n = 3). Scale bar represents 100 mm. (D and E) Immunohistochemical staining of CD31 (D) and a-SMA (E) in skin wounds from the control group, WT T1D group, and GCN2KO group at 7 dpi (n = 3). Scale bars represent 500 mm (top) and 50 mm (bottom). (F) Flow cytometry analysis of CD206 expression in 3 or 7 dpi skin macrophages from the control group, WT T1D group, and GCN2KO group (n = 3). (G) Real-time PCR showing CD206, Il10, Tgfb1, and Vegfa expression levels in 3 or 7 dpi skin macrophages from the control group, WT T1D group, and GCN2KO group (n = 3). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S6.

    Journal: Cell reports

    Article Title: Downregulation of nutrition sensor GCN2 in macrophages contributes to poor wound healing in diabetes.

    doi: 10.1016/j.celrep.2023.113658

    Figure Lengend Snippet: Figure 6. GCN2 is involved in T1D mouse delayed skin wound healing (A) Wound-healing pictures from control, T1D WT, and GCN2KO mice. (B) Wound-healing rates in the control group, WT T1D group, and GCN2KO group (n = 5). (C) Skin tissue H&E staining from the control group, WT T1D group, and GCN2KO group at 7 dpi (n = 3). Scale bar represents 100 mm. (D and E) Immunohistochemical staining of CD31 (D) and a-SMA (E) in skin wounds from the control group, WT T1D group, and GCN2KO group at 7 dpi (n = 3). Scale bars represent 500 mm (top) and 50 mm (bottom). (F) Flow cytometry analysis of CD206 expression in 3 or 7 dpi skin macrophages from the control group, WT T1D group, and GCN2KO group (n = 3). (G) Real-time PCR showing CD206, Il10, Tgfb1, and Vegfa expression levels in 3 or 7 dpi skin macrophages from the control group, WT T1D group, and GCN2KO group (n = 3). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S6.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies FITC anti-mouse CD206 BioLegend Cat# 141703; RRID: AB_10900988 PE anti-mouse F4/80 BioLegend Cat# 123110; RRID: AB_893486 APC anti-mouse F4/80 Thermo Fisher Cat# 17-4801-82; RRID: AB_2784648 PE/Cy5 anti-mouse CD11b Thermo Fisher Cat# 15-0112-83; RRID: AB_468714 Bv510 anti-mouse CD11b BioLegend Cat# 101263; RRID: AB_2629529 PE anti-mouse CD124 BioLegend Cat# 144804; RRID: AB_2561730 FITC anti-mouse F4/80 Thermo Fisher Cat# 11-4801-82; RRID: AB_2637191 PE anti-mouse TNFa Thermo Fisher Cat# 12-7321-82; RRID: AB_466199 FITC anti-mouse CD86 eBioscience Cat# 105006; RRID: AB_313149 PE/Cy5 anti-mouse CD45 Thermo Fisher Cat# 15-0459-42; RRID: AB_10717815 APC anti-mouse CD45 BioLegend Cat# 103112; RRID: AB_312977 PE anti-mouse Ly6G BioLegend Cat# 127607; RRID: AB_1186104 PE anti-mouse CD115 BioLegend Cat# 135506; RRID: AB_1937253 PE/Cy7 anti-mouse CD45.1 BioLegend Cat# 110730; RRID: AB_1134168 Fixable Viability Dye eFluorTM 780 Thermo Fisher Cat# 65-0865-14; RRID: —— PERCP/CY5.5 anti-mouse CD45.2 BioLegend Cat# 109827; RRID: AB_893352 Brilliant Violet 510 anti-mouse CD11b BioLegend Cat# 101245; RRID: AB_2561390 Brilliant Violet 605 anti-mouse Ly6C BioLegend Cat# 128036; RRID: AB_2562353 b-actin Sigma Cat# A5441; RRID: AB_476744 Arginase1 Santa Cruz Cat# sc-20150; RRID: AB_2058955 GCN2 Cell Signaling Technology Cat# 3302s; RRID: AB_10694800 p-IkBa Cell Signaling Technology Cat# 92456s; RRID: AB_2267145 IkBa Abcam Cat# ab32518; RRID: AB_2801653 p-IKKa/b Cell Signaling Technology Cat# 14938; RRID: AB_2798653 IKKa Cell Signaling Technology Cat# 61294s; RRID: AB_2799606 P65 Cell Signaling Technology Cat# 6596s; RRID: AB_2128043 p-P65 Cell Signaling Technology Cat# 3033T; RRID: AB_331284 Hif1a Cell Signaling Technology Cat# 36169; RRID: AB_2799095 p-eIF2a Cell Signaling Technology Cat# 9721s; RRID: AB_330951 eIF2a Cell Signaling Technology Cat# 9722s; RRID: AB_2230924 p-GCN2 Abcam Cat# ab75836; RRID: AB_1310260 Chemicals, peptides, and recombinant proteins Recombinant Murine M-CSF Pepro Tech Cat# 315-02 Thioglycollate BD BBL Cat# 211716 Recombinant murine IL4 R&D Systems Cat# 404-ML Trizol Thermo Fisher Cat# 15596018 Recombinant murine IL10 R&D Systems Cat# 417-ML Recombinant murine IFN-g R&D Systems Cat# 485-ML Lipopolysaccharide Sigma Cat# L2630 SYBR Green Mixture TaKaRa Cat# RR420A AMV Reverse transcriptase TaKaRa Cat# 2621 Oligo dT18 Thermo Fisher Cat# 18418020 Recombinant RNase Inhibitor TaKaRa Cat# 2313A (Continued on next page) 16 Cell Reports 43, 113658, January 23, 2024

    Techniques: Control, Staining, Immunohistochemical staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

    Figure 7. GCN2 agonist halofuginone (HF) rescued T1D mouse delayed wound healing (A) Immunoblot analysis showing GCN2, eIF2a, and their phosphorylated versions in skin wound macrophages from control and T1D mice treated with or without HF (n = 3). (B) Wound-healing pictures from control and T1D mice treated with or without HF. (C) Wound-healing rates in control and T1D mice treated with or without HF (n = 5). (D) Skin tissue H&E staining from control and T1D mice treated with or without HF at 7 dpi (n = 3). Scale bar represents 100 mm. (E and F) Immunohistochemical staining of CD31 (E) and a-SMA (F) in skin wounds from control and T1D mice treated with or without HF at 7 dpi (n = 3). Scale bars represent 500 mm (top) and 50 mm (bottom). (G) Flow cytometry analysis of CD206 expression in skin wound macrophages from control and T1D mice treated with or without HF at 7 dpi (n = 3). (H) Real-time PCR showing CD206, Il10, Tgfb1, and Vegfa expression levels in skin macrophages from control and T1D mice treated with or without HF at 3 or 7 dpi (n = 3). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S7.

    Journal: Cell reports

    Article Title: Downregulation of nutrition sensor GCN2 in macrophages contributes to poor wound healing in diabetes.

    doi: 10.1016/j.celrep.2023.113658

    Figure Lengend Snippet: Figure 7. GCN2 agonist halofuginone (HF) rescued T1D mouse delayed wound healing (A) Immunoblot analysis showing GCN2, eIF2a, and their phosphorylated versions in skin wound macrophages from control and T1D mice treated with or without HF (n = 3). (B) Wound-healing pictures from control and T1D mice treated with or without HF. (C) Wound-healing rates in control and T1D mice treated with or without HF (n = 5). (D) Skin tissue H&E staining from control and T1D mice treated with or without HF at 7 dpi (n = 3). Scale bar represents 100 mm. (E and F) Immunohistochemical staining of CD31 (E) and a-SMA (F) in skin wounds from control and T1D mice treated with or without HF at 7 dpi (n = 3). Scale bars represent 500 mm (top) and 50 mm (bottom). (G) Flow cytometry analysis of CD206 expression in skin wound macrophages from control and T1D mice treated with or without HF at 7 dpi (n = 3). (H) Real-time PCR showing CD206, Il10, Tgfb1, and Vegfa expression levels in skin macrophages from control and T1D mice treated with or without HF at 3 or 7 dpi (n = 3). Data represent three independent experiments and are presented as mean ± SD. Each independent experiment consists of at least three technical replicates. **p < 0.01, ***p < 0.001, one-way ANOVA. See also Figure S7.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies FITC anti-mouse CD206 BioLegend Cat# 141703; RRID: AB_10900988 PE anti-mouse F4/80 BioLegend Cat# 123110; RRID: AB_893486 APC anti-mouse F4/80 Thermo Fisher Cat# 17-4801-82; RRID: AB_2784648 PE/Cy5 anti-mouse CD11b Thermo Fisher Cat# 15-0112-83; RRID: AB_468714 Bv510 anti-mouse CD11b BioLegend Cat# 101263; RRID: AB_2629529 PE anti-mouse CD124 BioLegend Cat# 144804; RRID: AB_2561730 FITC anti-mouse F4/80 Thermo Fisher Cat# 11-4801-82; RRID: AB_2637191 PE anti-mouse TNFa Thermo Fisher Cat# 12-7321-82; RRID: AB_466199 FITC anti-mouse CD86 eBioscience Cat# 105006; RRID: AB_313149 PE/Cy5 anti-mouse CD45 Thermo Fisher Cat# 15-0459-42; RRID: AB_10717815 APC anti-mouse CD45 BioLegend Cat# 103112; RRID: AB_312977 PE anti-mouse Ly6G BioLegend Cat# 127607; RRID: AB_1186104 PE anti-mouse CD115 BioLegend Cat# 135506; RRID: AB_1937253 PE/Cy7 anti-mouse CD45.1 BioLegend Cat# 110730; RRID: AB_1134168 Fixable Viability Dye eFluorTM 780 Thermo Fisher Cat# 65-0865-14; RRID: —— PERCP/CY5.5 anti-mouse CD45.2 BioLegend Cat# 109827; RRID: AB_893352 Brilliant Violet 510 anti-mouse CD11b BioLegend Cat# 101245; RRID: AB_2561390 Brilliant Violet 605 anti-mouse Ly6C BioLegend Cat# 128036; RRID: AB_2562353 b-actin Sigma Cat# A5441; RRID: AB_476744 Arginase1 Santa Cruz Cat# sc-20150; RRID: AB_2058955 GCN2 Cell Signaling Technology Cat# 3302s; RRID: AB_10694800 p-IkBa Cell Signaling Technology Cat# 92456s; RRID: AB_2267145 IkBa Abcam Cat# ab32518; RRID: AB_2801653 p-IKKa/b Cell Signaling Technology Cat# 14938; RRID: AB_2798653 IKKa Cell Signaling Technology Cat# 61294s; RRID: AB_2799606 P65 Cell Signaling Technology Cat# 6596s; RRID: AB_2128043 p-P65 Cell Signaling Technology Cat# 3033T; RRID: AB_331284 Hif1a Cell Signaling Technology Cat# 36169; RRID: AB_2799095 p-eIF2a Cell Signaling Technology Cat# 9721s; RRID: AB_330951 eIF2a Cell Signaling Technology Cat# 9722s; RRID: AB_2230924 p-GCN2 Abcam Cat# ab75836; RRID: AB_1310260 Chemicals, peptides, and recombinant proteins Recombinant Murine M-CSF Pepro Tech Cat# 315-02 Thioglycollate BD BBL Cat# 211716 Recombinant murine IL4 R&D Systems Cat# 404-ML Trizol Thermo Fisher Cat# 15596018 Recombinant murine IL10 R&D Systems Cat# 417-ML Recombinant murine IFN-g R&D Systems Cat# 485-ML Lipopolysaccharide Sigma Cat# L2630 SYBR Green Mixture TaKaRa Cat# RR420A AMV Reverse transcriptase TaKaRa Cat# 2621 Oligo dT18 Thermo Fisher Cat# 18418020 Recombinant RNase Inhibitor TaKaRa Cat# 2313A (Continued on next page) 16 Cell Reports 43, 113658, January 23, 2024

    Techniques: Western Blot, Control, Staining, Immunohistochemical staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction