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cls-crown-like-structure

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HIF-2α is elevated in the ATMs from <em>db/db</em> mice. <span class="search-highlight">CLS</span>, <span class="search-highlight">crown</span>...
Published: 15 September 2014
HIF-2α is elevated in the ATMs from db/db mice. CLS, crown-like structure. HIF-2α is elevated in the ATMs from db/db mice. CLS, crown-like structure. More
Images
Representative Western blot analysis of ITCH protein in adipose tissue (AT)...
Published: 16 January 2014
Figure 7 Representative Western blot analysis of ITCH protein in adipose tissue (AT), isolated adipocytes (adipocyte fraction [AF]), and SVFs (A). Representative Western blot analysis of ITCH protein in AT, isolated adipocytes (AF), and SVFs (B). Images are representative of adipose tissue sections collected from five subjects. CLS, crown-like structure; ma, macrophage. The arrows indicate ITCH- and CD3-positive cells. Correlation between Itch and CD-206 mRNA (C) expression (r = −0.36, P = 0.01) and CD206-to-CD68 ratio (D) (r = −0.42, P = 0.003) in human adipose tissue (n = 50). Expression of mRNA was determined by real-time PCR and normalized to cyclophilin A. R.U., relative units; VAT, visceral adipose tissue. Figure 7. Representative Western blot analysis of ITCH protein in adipose tissue (AT), isolated adipocytes (adipocyte fraction [AF]), and SVFs (A). Representative Western blot analysis of ITCH protein in AT, isolated adipocytes (AF), and SVFs (B). Images are representative of adipose tissue sections collected from five subjects. CLS, crown-like structure; ma, macrophage. The arrows indicate ITCH- and CD3-positive cells. Correlation between Itch and CD-206 mRNA (C) expression (r = −0.36, P = 0.01) and CD206-to-CD68 ratio (D) (r = −0.42, P = 0.003) in human adipose tissue (n = 50). Expression of mRNA was determined by real-time PCR and normalized to cyclophilin A. R.U., relative units; VAT, visceral adipose tissue. More
Images
p38α in T cells promotes adipose tissue inflammation. WT and p38α<sup>ΔT</sup>...
Published: 29 March 2022
Figure 2 p38α in T cells promotes adipose tissue inflammation. WT and p38αΔT male mice were fed with HFD for 16 weeks. A: eWAT mass was measured. B: Representative H&E staining of eWAT, inguinal white adipose tissue (iWAT), and BAT sections (scale bars 100 μm). C: Immunohistochemistry of F4/80 in eWAT (scale bars 100 μm) (F4/80 is in brown) (left); number of crown-like structures was quantified (right). D: Quantification of CD4+ and CD8+ T-cell percentages and cell numbers in eWAT. E: Flow cytometric analysis of T-cell subset percentages in the eWAT of obese WT and p38αΔT male mice. F: Cell numbers of Th1, Th2, Th17, Treg, and TNF-α+ CD4+ and CD8+ T cells and IFN-γ+ CD8+ T cells in eWAT. G: Real-time PCR analysis of mRNA expression of indicated genes (normalized with Hprt) in eWAT. H: IFN-γ, IL-1β, IL-6, and TNF-α levels in the serum and eWAT homogenates of obese WT and p38αΔT male mice were measured by ELISA. Data are representative of two (CH) or three (A and B) independent experiments with four to six mice per group. Data are presented as mean ± SD. *P < 0.05; **P < 0.01. CLS, crown-like structures; N/A, not applicable; ns, not significant. More
Images
Effects of FO treatment on adipose macrophages and capillaries. Before and ...
Published: 16 April 2013
FIG. 2. Effects of FO treatment on adipose macrophages and capillaries. Before and after treatment with FO, adipose tissue from biopsies was analyzed histochemically. A: CD68 staining. A representative image showing macrophages (small arrows) and crown-like structures (large arrow). Data are expressed as mean ± SEM. B: Effects of FO and placebo on macrophage number (*P < 0.05 vs. pretreatment). C: Effects of treatment on the number of crown-like structures (CLS) (*P < 0.05 vs. pretreatment). D: Capillary and large vessels were identified by staining with lectin and α-smooth muscle actin, and representative images are shown. E: Effects of placebo and FO on the number of capillaries in adipose tissue. FIG. 2. Effects of FO treatment on adipose macrophages and capillaries. Before and after treatment with FO, adipose tissue from biopsies was analyzed histochemically. A: CD68 staining. A representative image showing macrophages (small arrows) and crown-like structures (large arrow). Data are expressed as mean ± SEM. B: Effects of FO and placebo on macrophage number (*P < 0.05 vs. pretreatment). C: Effects of treatment on the number of crown-like structures (CLS) (*P < 0.05 vs. pretreatment). D: Capillary and large vessels were identified by staining with lectin and α-smooth muscle actin, and representative images are shown. E: Effects of placebo and FO on the number of capillaries in adipose tissue. More
Meeting Abstracts
Journal: Diabetes
Diabetes 2020;69(Supplement_1):1680-P
Published: 01 June 2020
...) inflammation was decreased in ALA+CL compared to other groups, with lower serum IL-6 and IL-17 and lesser crown-like structures (CLS) on IHC for F4/80 and CD11c (M1-MΦ). Both IHC staining and mRNA expression of CD206 (M2-MΦ marker) confirm increased M2 MΦ’s in ALA+CL compared to other groups. Food intake...
Meeting Abstracts
Journal: Diabetes
Diabetes 2019;68(Supplement_1):306-LB
Published: 01 June 2019
... diet (HFD) induced obesity, macrophages, organized into crown like structures (CLS) around dead and dying adipocytes, are detected in the vicinity of type 1 adipocytes, as assessed by kernel density estimation. This association between type 1 adipocytes and CLS is secondary to a ∼10-fold increase...
Meeting Abstracts
Journal: Diabetes
Diabetes 2018;67(Supplement_1):2021-P
Published: 01 July 2018
...- Lpl, Fasn, Acsl1, Hsl, and Glut4 expression in obese females. Lipolysis promoted appearance of crown like structures (CLS) in males and females but only induced Il6 proinflammatory cytokine and Mcp1 chemokine expression in obese female GWAT. Activated lipolysis...
Meeting Abstracts
Journal: Diabetes
Diabetes 2018;67(Supplement_1):1877-P
Published: 01 July 2018
... mice in both strains, with no difference in weight gain between strains. Crown-like structure (CLS) number, a hallmark of AT inflammation, was increased in HV mice of both strains compared to CV mice. In addition, HV B6 mice had higher CLS number than HV B6129 mice. In B6129 mice, niacin decreased CLS...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(11):2802–2809
Published: 17 October 2011
... by higher secretion of proinflammatory cytokines and macrophage recruitment. Previous research has demonstrated that in adipose tissue from obese mice and humans, such macrophages aggregate around dead adipocytes, forming characteristic ring patterns referred to as crown-like structures (CLS) ( 8...
Journal Articles
Journal: Diabetes
Diabetes 2014;63(10):3161–3162
Published: 15 September 2014
...HIF-2α is elevated in the ATMs from db/db mice. CLS, crown-like structure. HIF-2α is elevated in the ATMs from db/db mice. CLS, crown-like structure. ...
Meeting Abstracts
Journal: Diabetes
Diabetes 2018;67(Supplement_1):1989-P
Published: 01 July 2018
... of eWAT in DIO mice revealed that CX3CR1 was predominantly expressed by F4/80+ macrophages in crown-like structures (CLS). On an HFD, KO mice had increased macrophage infiltration and formation of CLS in eWAT compared with those of WT mice, despite that weight and adipocyte size were similar...
Meeting Abstracts
Journal: Diabetes
Diabetes 2021;70(Supplement_1):257-OR
Published: 01 June 2021
..., 126±11 vs. 51±7 nmol/ml, p<0.005; phosphatidylcholine, 385±24 vs. 306±25 nmol/ml, p<0.01; ceramide, 1.7±0.2 vs. 1.2±0.2 nmol/ml, p<0.01) also increased after 12-hour glucagon infusion. H&E and CD68 immunofluorescence staining in adipose tissue displayed the crown-like structures (CLS...
Journal Articles
Journal: Diabetes
Diabetes 2016;65(12):3649–3659
Published: 13 September 2016
... neoangiogenesis via vascular endothelial growth factor (VEGF)-A. We previously reported that macrophages in crown-like structures (CLSs) are both hypoxic and inflammatory. In the current study, we examined how macrophage HIF-1α is involved in high-fat diet (HFD)–induced inflammation, neovascularization, hypoxia...
Includes: Supplementary data
Images
ATM and T-cell quantification in HFD-fed mice deficient for 5-LO or treated...
Published: 17 August 2012
FIG. 4. ATM and T-cell quantification in HFD-fed mice deficient for 5-LO or treated with the 5-LO inhibitor Zileuton. Immunohistochemical stainings for the macrophage marker MAC2 (A) or the T-cell marker CD3 (B) in paraffin-embedded epi-WAT sections from HFD-fed WT and 5-LO−/− mice and HFD-fed WT mice treated with vehicle or Zileuton. Quantitative representations of the illustrations on the left are shown in the graphs on the right of each panel, where ATM or T-cell presence is presented as crown-like structures (CLS) per 1,000 nuclei. qPCR analysis of relative mRNA levels of the macrophage marker CD68 (C) or the T-cell marker CD3 (E) in the same set of epi-WAT tissues. FACS analysis of SVCs derived from again the same samples, quantifying the total number of macrophages (D) or T cells (F) per gram epi-WAT and their composition in percentages of M2 (F4/80+CD11b+CD11c) and M1 (F4/80+CD11b+CD11c+) ATMs or CD4+ (CD3+CD4+CD8) and CD8+ (CD3+CD4CD8+) T cells, respectively. Data are expressed as mean ± SEM (n ≥ 6/group). *P < 0.05 and **P < 0.01 compared with WT or vehicle control groups. (A high-quality digital representation of this figure is available in the online issue.) FIG. 4. ATM and T-cell quantification in HFD-fed mice deficient for 5-LO or treated with the 5-LO inhibitor Zileuton. Immunohistochemical stainings for the macrophage marker MAC2 (A) or the T-cell marker CD3 (B) in paraffin-embedded epi-WAT sections from HFD-fed WT and 5-LO−/− mice and HFD-fed WT mice treated with vehicle or Zileuton. Quantitative representations of the illustrations on the left are shown in the graphs on the right of each panel, where ATM or T-cell presence is presented as crown-like structures (CLS) per 1,000 nuclei. qPCR analysis of relative mRNA levels of the macrophage marker CD68 (C) or the T-cell marker CD3 (E) in the same set of epi-WAT tissues. FACS analysis of SVCs derived from again the same samples, quantifying the total number of macrophages (D) or T cells (F) per gram epi-WAT and their composition in percentages of M2 (F4/80+CD11b+CD11c−) and M1 (F4/80+CD11b+CD11c+) ATMs or CD4+ (CD3+CD4+CD8−) and CD8+ (CD3+CD4−CD8+) T cells, respectively. Data are expressed as mean ± SEM (n ≥ 6/group). *P < 0.05 and **P < 0.01 compared with WT or vehicle control groups. (A high-quality digital representation of this figure is available in the online issue.) More
Images
MBL affects the development of diet-induced obesity in mice. <em>A</em>...
Published: 13 November 2014
Figure 1 MBL affects the development of diet-induced obesity in mice. A: Systemic levels of MBL-A after 16 weeks of following either an LFD or an HFD. B: Systemic levels of MBL-C after 16 weeks of following either an LFD or an HFD. C: Body weight development of MBL−/− and WT mice receiving an LFD or an HFD for 20 weeks (*P = 0.002 after 20 weeks). D: Energy expenditure was measured using indirect calorimetry. E: Representative H-E staining of liver sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×20). F: Representative images with F4/80+ staining to visualize Kupffer cells in liver (original magnification ×20). G: Quantitative determination of liver triglycerides (TG) in all four conditions after 20 weeks of the intervention. H: Representative H-E staining of WAT sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×20). I: Representative photographs of F4/80 staining in WAT sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×5 and ×20). Arrows indicate the presence of macrophages. J: Quantification of mean adipocyte size by determining the cross-sectional diameter for each condition (n = 5). K: Quantification of the number of crown-like structures (CLS) present in epididymal adipose tissue of WT and MBL−/− animals fed an HFD. *Significant difference at P < 0.05. Values are expressed as mean ± SEM (n = 11–13 animals in each group). For indirect calorimetry measurements in animals fed an HFD, six or seven animals per group were used. Figure 1. MBL affects the development of diet-induced obesity in mice. A: Systemic levels of MBL-A after 16 weeks of following either an LFD or an HFD. B: Systemic levels of MBL-C after 16 weeks of following either an LFD or an HFD. C: Body weight development of MBL−/− and WT mice receiving an LFD or an HFD for 20 weeks (*P = 0.002 after 20 weeks). D: Energy expenditure was measured using indirect calorimetry. E: Representative H-E staining of liver sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×20). F: Representative images with F4/80+ staining to visualize Kupffer cells in liver (original magnification ×20). G: Quantitative determination of liver triglycerides (TG) in all four conditions after 20 weeks of the intervention. H: Representative H-E staining of WAT sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×20). I: Representative photographs of F4/80 staining in WAT sections of MBL−/− and WT mice receiving an LFD or an HFD (original magnification ×5 and ×20). Arrows indicate the presence of macrophages. J: Quantification of mean adipocyte size by determining the cross-sectional diameter for each condition (n = 5). K: Quantification of the number of crown-like structures (CLS) present in epididymal adipose tissue of WT and MBL−/− animals fed an HFD. *Significant difference at P < 0.05. Values are expressed as mean ± SEM (n = 11–13 animals in each group). For indirect calorimetry measurements in animals fed an HFD, six or seven animals per group were used. More
Images
<em>Rnf20</em><sup>+/−</sup> mice exhibit reduced adiposity and mit...
Published: 11 October 2019
Figure 2 Rnf20+/− mice exhibit reduced adiposity and mitigated systemic insulin resistance upon HFD. A: Body weight change in WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8). B: Fat mass of WT and Rnf20+/− mice fed HFD for 7 weeks (n = 3). Fat mass was measured by 1H MRS. C: Representative whole-mount BODIPY staining image of eWAT from WT and Rnf20+/− mice fed HFD for 13 weeks. D: Adipocyte size of eWAT from WT and Rnf20+/− mice fed HFD for 5, 10, and 18 weeks (w). Size of 250–500 adipocytes was measured in each group. E: Representative hematoxylin-eosin (H&E) staining (left) and whole-mount immunocytochemistry image (right) of eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks. F: Number of crown-like structures (CLS) in eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8). G: qRT-PCR analyses of proinflammatory genes in eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks (n = 4–8). HN: Serum profiles and basal glycerol release of WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8; n = 3 in J): serum FFA (H), fasting serum triglyceride (TG) (I), fasting serum cholesterol (J), basal glycerol release of primary adipocytes of WT and Rnf20+/− mice (K), fasting blood glucose (L), fasting serum insulin (M), and HOMA-IR (N). O: Intraperitoneal GTT and area under the curve (AUC) (n = 6–8). P: Intraperitoneal ITT (n = 6–8). Data are mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Ct, cycle threshold; n.s. nonsignificant. Figure 2. Rnf20+/− mice exhibit reduced adiposity and mitigated systemic insulin resistance upon HFD. A: Body weight change in WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8). B: Fat mass of WT and Rnf20+/− mice fed HFD for 7 weeks (n = 3). Fat mass was measured by 1H MRS. C: Representative whole-mount BODIPY staining image of eWAT from WT and Rnf20+/− mice fed HFD for 13 weeks. D: Adipocyte size of eWAT from WT and Rnf20+/− mice fed HFD for 5, 10, and 18 weeks (w). Size of 250–500 adipocytes was measured in each group. E: Representative hematoxylin-eosin (H&E) staining (left) and whole-mount immunocytochemistry image (right) of eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks. F: Number of crown-like structures (CLS) in eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8). G: qRT-PCR analyses of proinflammatory genes in eWAT from WT and Rnf20+/− mice fed HFD for 18 weeks (n = 4–8). H–N: Serum profiles and basal glycerol release of WT and Rnf20+/− mice fed HFD for 18 weeks (n = 6–8; n = 3 in J): serum FFA (H), fasting serum triglyceride (TG) (I), fasting serum cholesterol (J), basal glycerol release of primary adipocytes of WT and Rnf20+/− mice (K), fasting blood glucose (L), fasting serum insulin (M), and HOMA-IR (N). O: Intraperitoneal GTT and area under the curve (AUC) (n = 6–8). P: Intraperitoneal ITT (n = 6–8). Data are mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Ct, cycle threshold; n.s. nonsignificant. More
Images
Hematopoietic deletion of CD36 reduces macrophage content and improves insu...
Published: 22 March 2011
FIG. 2. Hematopoietic deletion of CD36 reduces macrophage content and improves insulin signaling in adipose tissue of mice fed an HFD. Mice with hematopoietic-specific deletion of CD36 (CD36KO) were created by bone marrow transplanting stem cells from CD36KO or background-matched wild-type (WT) mice into lethally irradiated C57BL/6 mice. After 20 weeks of the standard diet or HFD, epididymal WAT depots were excised before and after insulin stimulation. A: Gene expression of F4/80 (Emr1), CD11c (Itgax), and ccl2 in the WAT of WT and CD36KO mice on an HFD was measured by real-time PCR, expressed relative to WT standard diet–fed mice indicated by the dotted line. B: Immunohistochemical staining for F4/80 (brown staining, see arrow) in WAT sections from WT and CD36KO mice on an HFD. The percentage of adipocytes with crown-like structures (CLS) was determined for both WT and CD36KO mice on an HFD (6–10 images at ×20 magnification were analyzed per mouse). C: Gene expression of the alternative macrophage activation (M2) marker, arginase 1 (Arg1), and inflammatory cytokines tnfa, il6, and ifng in the WAT of WT and CD36KO mice were measured by real-time PCR, expressed relative to WT standard diet–fed mice, indicated by the dotted line. WAT from basal and insulin-stimulated WT and CD36KO mice fed a standard diet and HFD were analyzed for phosphorylated (p) (Tyr1361) and total IR (D) and phosphorylated (p) (Ser473 and Thr308) and total Akt (E and F) by Western blotting. Data are expressed as means ± SE (n = 6–8 mice per group). *P < 0.05 insulin stimulated vs. basal; **P < 0.001 insulin stimulated vs. basal; #P < 0.05 CD36KO vs. WT; ##P < 0.001 CD36KT; Student t tests and two-way ANOVA with post hoc analysis. (A high-quality digital representation of this figure is available in the online issue.) FIG. 2. Hematopoietic deletion of CD36 reduces macrophage content and improves insulin signaling in adipose tissue of mice fed an HFD. Mice with hematopoietic-specific deletion of CD36 (CD36KO) were created by bone marrow transplanting stem cells from CD36KO or background-matched wild-type (WT) mice into lethally irradiated C57BL/6 mice. After 20 weeks of the standard diet or HFD, epididymal WAT depots were excised before and after insulin stimulation. A: Gene expression of F4/80 (Emr1), CD11c (Itgax), and ccl2 in the WAT of WT and CD36KO mice on an HFD was measured by real-time PCR, expressed relative to WT standard diet–fed mice indicated by the dotted line. B: Immunohistochemical staining for F4/80 (brown staining, see arrow) in WAT sections from WT and CD36KO mice on an HFD. The percentage of adipocytes with crown-like structures (CLS) was determined for both WT and CD36KO mice on an HFD (6–10 images at ×20 magnification were analyzed per mouse). C: Gene expression of the alternative macrophage activation (M2) marker, arginase 1 (Arg1), and inflammatory cytokines tnfa, il6, and ifng in the WAT of WT and CD36KO mice were measured by real-time PCR, expressed relative to WT standard diet–fed mice, indicated by the dotted line. WAT from basal and insulin-stimulated WT and CD36KO mice fed a standard diet and HFD were analyzed for phosphorylated (p) (Tyr1361) and total IR (D) and phosphorylated (p) (Ser473 and Thr308) and total Akt (E and F) by Western blotting. Data are expressed as means ± SE (n = 6–8 mice per group). *P < 0.05 insulin stimulated vs. basal; **P < 0.001 insulin stimulated vs. basal; #P < 0.05 CD36KO vs. WT; ##P < 0.001 CD36KT; Student t tests and two-way ANOVA with post hoc analysis. (A high-quality digital representation of this figure is available in the online issue.) More
Journal Articles
Journal: Diabetes
Diabetes 2007;56(12):2910–2918
Published: 01 December 2007
...@tufts.edu 27 8 2007 5 6 2007 DIABETES 2007 AT, adipose tissue ATMΦ, AT macrophage AUC, area under the curve CCR, C-C chemokine receptor CLS, crown-like structure DC, dendritic cell DIO, diet-induced obesity eAT, epididymal AT HF, high fat iAT, inguinal subcutaneous AT IL...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2014;63(2):550–561
Published: 16 January 2014
... of adipose tissue sections collected from five subjects. CLS, crown-like structure; ma, macrophage. The arrows indicate ITCH- and CD3-positive cells. Correlation between Itch and CD-206 mRNA (C) expression (r = −0.36, P = 0.01) and CD206-to-CD68 ratio (D...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2007;56(6):1517–1526
Published: 01 June 2007
...+ lectin-binding cells could clearly be distinguished from CD34 CD68+ macrophages, which were scattered in the stroma and did not bind lectin. Adipogenic/angiogenic cell clusters can morphologically and immunohistochemically be distinguished from crown-like structures frequently seen...
Includes: Multimedia, Supplementary data