Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: ( A ) GO analysis of macrophage response to LNR. ( B ) Heatmaps of macrophage-secreted chemokines in the iWAT from mice receiving sham or LNR at 6°C for 7 days ( n = 4 per group). Red and blue represent the fold increase and decrease in a gene, respectively (see color scale). ( C ) mRNA expression of macrophage-secreted chemokines in iWAT from mice described in (B) ( n = 8 per group). ( D ) Ccl22 mRNA expression in iWAT from mice described in (B) at 23° or 6°C for 7 days ( n = 6 per group). ( E ) Ccl22 mRNA expression in iWAT from mice described in (B) exposed to 6°C for different hours ( n = 6). ( F ) Serum CCL22 levels from mice described in (B) exposed to 6°C for different days ( n = 6). ( G ) Serum CCL22 levels from mice described in (B) ( n = 6 per group). ( H ) Ccl22 mRNA expression in iWAT SVF cells or mature adipocytes (MAs). ( I ) Ccl22 mRNA expression in M0, M1, and M2 macrophages derived from 10-week-old C57BL/6 bone marrows ( n = 8 per group). ( J and K ) mRNA expression (J) and IF (K) of UCP1 in beige adipocytes ( n = 6 per group). Scale bars, 25 μm. ( L to N ) Immunofluorescence of UCP1 (L), mRNA expression of thermogenic genes (M), and immunoblots of UCP1 (N) in iWAT ( n = 3 to 6 per group). Scale bars, 50 μm. ( O to R ) Immunoblots and quantification of UCP1 [(O) and (P)], mRNA expression (Q) of thermogenic genes, and IF of UCP1 and H&E staining (R) in iWAT ( n = 3 to 6 per group). Scale bars, 50 μm. Data information: Results are presented as means ± SEM. [(C), (D), (G) to (J), (M), (P), and (Q)] * P ≤ 0.05, ** P < 0.01, and *** P < 0.001 by nonpaired Student’s t test. [(E) and (F)] * P ≤ 0.05 and ** P < 0.01 nonpaired Student’s t test compared with before cold stimulation.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Expressing, Derivative Assay, Immunofluorescence, Western Blot, Staining

Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: ( A ) Pearson’s correlation coefficient analysis of serum CCL22 concentration and fat mass ( n = 28 per group). ( B to F ) Body weight (B), energy consumption rate [(C) and (D)], and RER [(E) and (F)] ( n = 8 per group). Ten-week-old C57BL/6 male mice received saline or rCCL22 (20 μg/kg per day) at 6°C for 14 days with HFD. ( G to I ) Immunofluorescence (G), immunoblots (H) of UCP1, and mRNA expression (I) of thermogenic genes in iWAT ( n = 3 to 6 per group). Scale bar, 50 μm. ( J to L ) Immunofluorescence (J), immunoblots, and quantification [(K) and (L)] of UCP1 in iWAT ( n = 3 per group). SVF cells were extracted from HFD-fed iWAT, cultured with vehicle or rCCL22 (10 ng/ml) for 4 days, and then induced to beige adipocytes for 5 days. Scale bars, 25 μm. ( M ) Schematic diagram: The 6-month trial of obese adults participated in ADF: a 3-month weight loss period followed by a 3-month weight maintenance period. ( N and O ) Pearson’s correlation coefficient analysis of human serum CCL22 levels, body weight (N), and fat mass (O) ( n = 40). ( P ) Human serum CCL22 levels ( n = 20). ( Q ) Schematic diagram: Human SVFs from iWAT were cultured at 31°C for 5 days, treated with vehicle or CCL22 (10 ng/ml) for 4 days, and then induced to beige adipocytes. ( R to T ) Immunoblots and quantification [(R) and (S)] of UCP1 and mRNA expression (T) of thermogenic genes in human beige adipocytes ( n = 3 to 6 per group). Data information: Results are presented as means ± SEM. [(A), (N), and (O)] Two-tailed Pearson’s correlation coefficient analysis. [(F), (I), (L), (S), and (T)] * P ≤ 0.05 and ** P < 0.01 by nonpaired Student’s t test. (P) ** P < 0.01 by nonpaired Student’s t test compared with month 0 body weight. (B) * P ≤ 0.05 by two-way analysis of variance (ANOVA) followed by post hoc Bonferroni tests.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Concentration Assay, Saline, Immunofluorescence, Western Blot, Expressing, Cell Culture, Two Tailed Test

Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: Primer sequences for qPCR gene expression analysis.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Gene Expression