proteoglycans mimecan  (R&D Systems)

Journal: Frontiers in cellular and infection microbiology

Article Title: Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity.

doi: 10.3389/fcimb.2022.966370

Figure Lengend Snippet: FIGURE 6 Proteolytic activity of L. interrogans leptolysin on proteoglycans and plasma fibronectin. (A) Degradation of substrates: mimecan, lumican, biglycan, decorin (0.5 mg), and plasma fibronectin (5 mg) were incubated with leptolysin (0.1 µg) at 37°C for the indicated time points. Cleavage products were subjected to SDS- polyacrylamide gel under reducing conditions, transferred to nitrocellulose membranes, and probed with specific antibodies, or the gel was silver stained (FP). (B) Inhibition of proteolytic activity: before the addition of each substrate, leptolysin (0.1 µg) was incubated with 5 mmol/L 1,10-phenanthroline (lane 4) for 30 min at room temperature. Substrates were added and incubations proceeded for 24 h. Cleavage products were analyzed as described above. Mim (mimecan), Lum (lumican), Bgn (biglycan), Dec (decorin), FP (plasma fibronectin), Sup (Leptospira culture supernatant), Leptol. (leptolysin), Fen (1,10-phenanthroline).

Article Snippet: Recombinant L. interrogans leptolysin (0.1 mg) was incubated with 0.5 mg of proteoglycans mimecan (# 2949-MC), lumican (# 2846-LU), biglycan (# 2667-CM) and decorin (# 143- DE) (R & D Systems, Inc., Minneapolis, USA), and with 5 μg of plasma fibronectin (Sigma-Aldrich, St. Louis, MO, USA) for up to 4 h in 50 mM Tris HCl, 200 mM NaCl, 10 mM CaCl2 and 0.05% CHAPS, pH 7.4, at 37°C.

Techniques: Activity Assay, Clinical Proteomics, Incubation, Staining, Inhibition

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoblastic Y1 receptor signaling modulates insulin secretion and insulin action via osteoglycin. Scatter plots showing osteoblast cells isolated from (A) non-GFP expressing mice and (B) GFP expressing mice showing GFP signal, forward scatter (FSC) and cell count. Green GFP positive (GFP POS) gate represents the purified osteoblast cells that are selected for further analysis. (C) Representative photomicrograph showing GFP positive osteoblasts. qPCR of osteoglycin mRNA levels in (D) wildtype (WT) and Y1 −/− osteoblastic cells and (E) WT and Y1 lox/lox 3.6Cre femora. (F) qPCR of Ins1 and Ins2 mRNA levels in MIN6 cells with different doses of osteoglycin. (G) Insulin secretion from MIN6 cells during a glucose stimulated insulin secretion test with or without osteoglycin (1 μg/mL). Data are means ± SEM of at least 6 per group, * = p < 0.05 as indicated. Scale bar represents 5 μm.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques: Isolation, Expressing, Cell Counting, Purification

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoglycin deletion affects glucose and insulin homeostasis. (A–B) Glucose and (C–D) insulin levels during a glucose tolerance test shown either as a time course (A, C) or expressed as area under the curve (B, D) for Ogn −/− and wildtype (WT) mice on both chow and high fat diet. (E) Glucose levels during an insulin tolerance test shown as a time course and (F) expressed as area under the curve for Ogn −/− and WT mice on both chow and high fat diet. (G) Serum glucose and (H) serum insulin levels in Ogn −/− and WT mice on both chow and high fat diet at cull at 16 weeks of age. Histological analysis showing (I) number of islets and (J) beta cell area in the pancreas of Ogn −/− and WT mice on chow diet. Data are means ± SEM of at least 10 mice per group, except 4–6 mice per group for islet histology (I–J). * = p < 0.05, ** = p < 0.01, *** = p < 0.001 between chow groups or as indicated; # = p < 0.05, ## = p < 0.01 between HFD groups.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques:

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoglycin treatment controls glucose and insulin homeostasis. (A) Glucose levels during a pilot glucose tolerance test in WT mice treated with either PBS or different doses of osteoglycin 4 h prior to the test. (B) Glucose and (C) insulin levels in fasted WT mice 4 h after treatment with either osteoglycin or PBS. (D) Glucose levels during a glucose tolerance test and (E) expressed as area under the curve in chow-fed WT mice as well as (F) glucose levels during a glucose tolerance test in HFD-fed WT mice treated with either osteoglycin or PBS 4 h prior to the test. (G) Insulin levels during the glucose tolerance test in chow-fed WT mice treated with either osteoglycin or PBS 4 h prior to the test. (H) Glucose levels during an insulin tolerance test in WT mice treated with either osteoglycin or PBS 4 h prior to the test. (I–J) pAkt (Ser473) in quadriceps muscle of WT mice treated with osteoglycin or PBS followed by either saline or insulin (0.5 IU/kg) 4 h later. (I) pAkt to Akt ratios of treatment relative to untreated group; (J) representative immunoblots. Data are means ± SEM of at least 10 mice per group, except >3 mice per group for pilot study (A), and 4–6 mice per group for pAkt experiment (I–J). * = p < 0.05, ** = p < 0.01 as indicated; ˆ = p < 0.05, ˆˆ = p < 0.01 OGN treatment effect over time compared to PBS as measured by repeated measures ANOVA.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques: Saline, Western Blot

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Lack of osteoglycin leads to an increase in bone mass. (A) BMD and (B) BMC of isolated femora from Ogn −/− and wildtype (WT) mice as measured by DXA. (C) Femur length. (D) Bone area as measured by micro-cT across the length of femora also represented as (E) area under the curve. Micro-cT analysis of femoral trabecular bone showing (F) BV/TV, (G) trabecular thickness, and (H) trabecular number. Micro-cT analysis of femoral cortical bone showing (I) bone volume, (J) cortical thickness, (K) periosteal perimeter, and (L) mean polar moment of inertia (MMI). (M) Representative sagittal and (N) cross-sectional images from femora of Ogn −/− and WT mice. Vertebral micro-cT analysis of trabecular bone in the 3rd lumbar vertebrae showing (O) BV/TV, (P) trabecular thickness, (Q) trabecular number and (R) representative images. Histomorphometric analysis showing (S) mineral apposition rate (MAR), (T) bone formation rate (BFR), (U) mineralising surface, (V) osteoclast number, and (W) osteoclast surface in the femora of Ogn −/− and WT mice. Data are means ± SEM of at least 5–7 per group, * = p < 0.05, ** = p < 0.01 as indicated.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques: Isolation, Micro-CT

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Lack of osteoglycin alters metabolic and activity parameters. (A) Averages for food intake during the dark and light phases, (B) cumulative food intake over a 72 h time course, (C) averages for distance in voluntary locomotion travelled (pedmeters) during the dark and light phases, (D) cumulative pedmeters over a 72 h time course, (E) averages for energy expenditure during the dark and light phases, (F) energy expenditure over a 72 h time course, (G) averages for respiratory quotient (RQ) during the dark and light phases, and (H) RQ over a 72 h time course in Ogn −/− compared to wildtype (WT) mice. Data are means ± SEM of at least 4 per group, * = p < 0.05 as indicated. Shaded regions represent dark phase.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques: Activity Assay

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoglycin modulates central NPY to alter food intake. Cumulative food intake in wildtype mice treated with either osteoglycin (OGN) or PBS control (A) prior to treatment and (B) immediately following treatment. (C) Total food intake during the first 6 h following OGN or PBS treatment. (D) Representative photomicrographs and (E) quantification showing cfos expression in the Arcuate nucleus of the hypothalamus following i.p. injections of either saline or OGN. (F) Representative photomicrographs and (G) quantification showing cfos (red), NPY (green) and cfos/NPY co-localization (yellow) expression in the Arcuate nucleus of the hypothalamus following icv injections of either saline or OGN. (H) qPCR of NPY mRNA levels in the Arcuate of WT and Ogn −/− brains. Data are means ± SEM of at least 4 per group, * = p < 0.05, ** = p < 0.01, *** = p < 0.001 as indicated. Scalebar represents 100 μm.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques: Control, Expressing, Saline

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoglycin negatively correlates with fat and bone mass in mice. (A) Osteoglycin levels at cull in wildtype (WT) mice fed either a chow or HFD. Correlations between osteoglycin levels and (B) fat mass, (C) glucose levels, (D) insulin levels and (E) whole body BMC as well as correlation between (F) glucose levels and whole body BMC in WT mice fed either chow or HFD. (G) Body weight and (H) total white adipose tissue (WAT) mass in WT and Ogn −/− mice fed either a chow or HFD for 8 weeks. uCT analysis of the femora of WT or Ogn −/− mice fed either a chow or HFD showing (I) cortical bone volume, (J) cortical thickness, (K) periosteal perimeter, (L) polar MMI, (M) trabecular bone volume (BV/TV), (N) trabecular thickness, and (O) trabecular number. Data are means ± SEM of at least 7 per group, * = p < 0.05, ** = p < 0.01, *** = p < 0.001 as indicated.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques:

Journal: Molecular Metabolism

Article Title: Osteoglycin, a novel coordinator of bone and glucose homeostasis

doi: 10.1016/j.molmet.2018.05.004

Figure Lengend Snippet: Osteoglycin response correlates with the glycemic response to weight loss in severely obese humans. Baseline correlations between osteoglycin levels and (A) body weight, (B) fat mass, (C) BMI, and (D) lean mass in severely obese patients in a weight loss intervention study. Correlations between the change in osteoglycin levels from pre-operative state to 1 month post-intervention with (E) bodyweight change, (F) BMI change and (G) glucose levels at 1 month post-intervention. Data are means ± SEM of at least 19 per group, p values are as indicated. NS = not significant.

Article Snippet: MIN6 cells were treated for 4 h with recombinant mouse osteoglycin (R&D Systems) at the stated doses after which RNA was extracted from the cells or the cells were subjected to a glucose stimulated insulin secretion (GSIS) test as described previously .

Techniques:

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Mimecan expressed in various mouse tissues and secreted into the blood. (a) Mimecan mRNA expression in various tissues from C57BL/6J mice determined by northern blot analysis. (b) Mimecan corresponding to 25 kDa and 12 kDa was detected in human serum by western blot analysis using monoclonal antibody against mimecan. H1–H4 represents four healthy individuals. M: molecular weight markers. Body mass index (BMI) of the individuals: H1, 21.8; H2, 21.5; H3, 19.5; H4, 25.7. (c) Northern blot analysis of mimecan mRNA expression in adipose tissue from C57BL/6J mice fasting for 0, 24, and 48 h (n = 4 for each time point). The right histogram is the gray scale of mimecan mRNA expression. Data are expressed as means ± SEM. * P < 0.05 vs 0 h. Statistical analysis was performed by one-way ANOVA.

Article Snippet: Moreover, both IL-1β and IL-6 were induced in the microglia by recombinant mouse mimecan (R&D, Minneapolis MN, USA), which was purified from a eukaryotic expression system.

Techniques: Expressing, Northern Blot, Western Blot, Molecular Weight

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) injections of mimecan induced a reduction in food intake and the anorexic effect of mimecan was attenuated by pretreatment with polyclonal antibody or by heat-inactivated mimecan. (a) Cumulative food intake of C57BL/6J mice responding to i.p. injection of different doses of mimecan-MBP (Mim-MBP) at indicated time points after 12 h of fasting (8:00 p.m.–8:00 a.m., n = 8). (b) Suppression of food intake in C57BL/6J mice with ad libitum access to food when responding to i.p. injection of Mim-MBP (0.05 μmol/kg) (n = 5), compared to phosphate-buffered saline (PBS) or maltose binding protein (MBP). (c) Suppression of food intake in Spraque–Dawley (SD) rats responding to i.c.v. injection of mimecan-MBP (Mim-MBP) (2 nmol/kg) (n = 8). (d) Cumulative food intake of C57BL/6J mice after i.p. injection with active or inactive Mim-MBP (n = 5). (e) Antibody neutralization test in C57BL/6J mice. Mice were pretreated with polyclonal antibody against mimecan or rabbit preimmune IgG by i.p. injection (n = 12). (f) Suppression of food intake of C57BL/6J mice responding to i.p. injection of mimecan-His (Mim-His) (0.05 μmol/kg) (n = 6). * P < 0.05; ** P < 0.01 vs. PBS at the indicated time points in (a) and (b). * P < 0.05; ** P < 0.01 vs. Mim-MBP inactivated at time points in (d). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: Moreover, both IL-1β and IL-6 were induced in the microglia by recombinant mouse mimecan (R&D, Minneapolis MN, USA), which was purified from a eukaryotic expression system.

Techniques: Injection, Saline, Binding Assay, Neutralization

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: The anorexic effect of mimecan was independent of leptin and melanocortin signaling. (a, b) Effect of mimecan on daily food intake (a) and body weight increment (b) in db / db mice with ad libitum access to food (0.05 μmol/kg; i.p. injection, n = 9–10). (b) Daily body weight change (g) = body weight on the indicated day — body weight on day 0. The solid circles and squares represent the changes of body weight during mimecan-MBP or MBP treatment, respectively. The empty circles and squares represent the changes of body weight after stopping injections of mimecan-MBP or MBP, respectively. (c) Cumulative food intake of A y /a mice treated with Mim-MBP (0.05 μmol/kg) after fasting for 6 h (i.p. injection, n = 4–5). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: Moreover, both IL-1β and IL-6 were induced in the microglia by recombinant mouse mimecan (R&D, Minneapolis MN, USA), which was purified from a eukaryotic expression system.

Techniques: Injection

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Effect of mimecan on IL-1β, IL-6, and SOCS3 expression and features of mimecan knockout mice. (a) The mRNA expression of IL-1β, IL-6, and SOCS3 in the hypothalamus from C57BL/6J mice (n = 10) with ad libitum access to food, treated with active or inactive Mim-MBP (0.05 μmol/kg) by i.p. injection for 4 h. The reference gene was actin. (b) Effect of recombinant mouse mimecan (rm Mim) (100 nM for 4 h) on the expression of IL-1β, SOCS3, and IL-6 in N9 microglia cells. The reference gene was actin. (c, d) The body weight (c) and daily food intake (d) in Min − / − (knockout) mice and wild type (WT) littermates with ad libitum access to food (n = 25). (e) Leptin mRNA expression in adipose tissue from Min − / − mice and WT littermates (n = 4). (f) Levels of leptin in serum from Mim − / − mice and WT littermates (n = 12). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: Moreover, both IL-1β and IL-6 were induced in the microglia by recombinant mouse mimecan (R&D, Minneapolis MN, USA), which was purified from a eukaryotic expression system.

Techniques: Expressing, Knock-Out, Injection, Recombinant

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Mimecan expressed in various mouse tissues and secreted into the blood. (a) Mimecan mRNA expression in various tissues from C57BL/6J mice determined by northern blot analysis. (b) Mimecan corresponding to 25 kDa and 12 kDa was detected in human serum by western blot analysis using monoclonal antibody against mimecan. H1–H4 represents four healthy individuals. M: molecular weight markers. Body mass index (BMI) of the individuals: H1, 21.8; H2, 21.5; H3, 19.5; H4, 25.7. (c) Northern blot analysis of mimecan mRNA expression in adipose tissue from C57BL/6J mice fasting for 0, 24, and 48 h (n = 4 for each time point). The right histogram is the gray scale of mimecan mRNA expression. Data are expressed as means ± SEM. * P < 0.05 vs 0 h. Statistical analysis was performed by one-way ANOVA.

Article Snippet: On day 7, cells were treated with 10, 50, or 100 nM recombinant mouse mimecan (R&D Systems) for 1, 2, or 4 h, and the expression of appetite-regulation neuropeptides and hypothalamic inflammatory factors was analyzed.

Techniques: Expressing, Northern Blot, Western Blot, Molecular Weight

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) injections of mimecan induced a reduction in food intake and the anorexic effect of mimecan was attenuated by pretreatment with polyclonal antibody or by heat-inactivated mimecan. (a) Cumulative food intake of C57BL/6J mice responding to i.p. injection of different doses of mimecan-MBP (Mim-MBP) at indicated time points after 12 h of fasting (8:00 p.m.–8:00 a.m., n = 8). (b) Suppression of food intake in C57BL/6J mice with ad libitum access to food when responding to i.p. injection of Mim-MBP (0.05 μmol/kg) (n = 5), compared to phosphate-buffered saline (PBS) or maltose binding protein (MBP). (c) Suppression of food intake in Spraque–Dawley (SD) rats responding to i.c.v. injection of mimecan-MBP (Mim-MBP) (2 nmol/kg) (n = 8). (d) Cumulative food intake of C57BL/6J mice after i.p. injection with active or inactive Mim-MBP (n = 5). (e) Antibody neutralization test in C57BL/6J mice. Mice were pretreated with polyclonal antibody against mimecan or rabbit preimmune IgG by i.p. injection (n = 12). (f) Suppression of food intake of C57BL/6J mice responding to i.p. injection of mimecan-His (Mim-His) (0.05 μmol/kg) (n = 6). * P < 0.05; ** P < 0.01 vs. PBS at the indicated time points in (a) and (b). * P < 0.05; ** P < 0.01 vs. Mim-MBP inactivated at time points in (d). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: On day 7, cells were treated with 10, 50, or 100 nM recombinant mouse mimecan (R&D Systems) for 1, 2, or 4 h, and the expression of appetite-regulation neuropeptides and hypothalamic inflammatory factors was analyzed.

Techniques: Injection, Saline, Binding Assay, Neutralization

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: The anorexic effect of mimecan was independent of leptin and melanocortin signaling. (a, b) Effect of mimecan on daily food intake (a) and body weight increment (b) in db / db mice with ad libitum access to food (0.05 μmol/kg; i.p. injection, n = 9–10). (b) Daily body weight change (g) = body weight on the indicated day — body weight on day 0. The solid circles and squares represent the changes of body weight during mimecan-MBP or MBP treatment, respectively. The empty circles and squares represent the changes of body weight after stopping injections of mimecan-MBP or MBP, respectively. (c) Cumulative food intake of A y /a mice treated with Mim-MBP (0.05 μmol/kg) after fasting for 6 h (i.p. injection, n = 4–5). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: On day 7, cells were treated with 10, 50, or 100 nM recombinant mouse mimecan (R&D Systems) for 1, 2, or 4 h, and the expression of appetite-regulation neuropeptides and hypothalamic inflammatory factors was analyzed.

Techniques: Injection

Journal: EBioMedicine

Article Title: Mimecan, a Hormone Abundantly Expressed in Adipose Tissue, Reduced Food Intake Independently of Leptin Signaling

doi: 10.1016/j.ebiom.2015.09.044

Figure Lengend Snippet: Effect of mimecan on IL-1β, IL-6, and SOCS3 expression and features of mimecan knockout mice. (a) The mRNA expression of IL-1β, IL-6, and SOCS3 in the hypothalamus from C57BL/6J mice (n = 10) with ad libitum access to food, treated with active or inactive Mim-MBP (0.05 μmol/kg) by i.p. injection for 4 h. The reference gene was actin. (b) Effect of recombinant mouse mimecan (rm Mim) (100 nM for 4 h) on the expression of IL-1β, SOCS3, and IL-6 in N9 microglia cells. The reference gene was actin. (c, d) The body weight (c) and daily food intake (d) in Min − / − (knockout) mice and wild type (WT) littermates with ad libitum access to food (n = 25). (e) Leptin mRNA expression in adipose tissue from Min − / − mice and WT littermates (n = 4). (f) Levels of leptin in serum from Mim − / − mice and WT littermates (n = 12). Data are expressed as means ± SEM. * P < 0.05; ** P < 0.01. Statistical analysis was performed by one-way ANOVA.

Article Snippet: On day 7, cells were treated with 10, 50, or 100 nM recombinant mouse mimecan (R&D Systems) for 1, 2, or 4 h, and the expression of appetite-regulation neuropeptides and hypothalamic inflammatory factors was analyzed.

Techniques: Expressing, Knock-Out, Injection, Recombinant