recombinant mouse il 10  (MedChemExpress)

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

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

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

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