Journal: International Journal of Molecular Sciences

Article Title: Estrogen Impairs Adipose Tissue Expansion and Cardiometabolic Profile in Obese-Diabetic Female Rats

doi: 10.3390/ijms222413573

Figure Lengend Snippet: Ovariectomy and E2 treatment effects on oral glucose and insulin tolerance tests in female ZDF rats. ( A ) Oral Glucose Tolerance Test (OGTT). ( B ) Insulin Tolerant Test (ITT). In A and B, left panels represent blood glucose curves and right panels represent AUC. Blood samples were obtained from the tail vein at indicated time points. ( C ) Ratio of pAKT/AKT and protein levels of GLUT4 in WAT. ( D ) Cd36 and Ap2 mRNA expression levels in WAT. AUC, area under the curve; AKT, protein kinase B; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GLUT4, glucose transporter type 4; Cd36 , cluster of differentiation 36; Ap2 , adaptor protein 2. In C pAKT protein bands were normalized to total AKT protein intensity, and GLUT4 protein bands were normalized to GAPDH loading control. mRNA levels were normalized to housekeeping Gapdh expression. In C and D, data are expressed in arbitrary units (au) related to LEAN group. Values are expressed as the mean ± SEM. Differences between obese-diabetic (SHAM) and control (LEAN) animals were analyzed by Student’s t -test. Differences between obese-diabetic groups with different hormonal manipulation were analyzed by One-way ANOVA with Fisher’s LSD post-hoc test. Student’s t -test: a, SHAM vs. LEAN. One-way ANOVA: b, OVA or OVA + E2 vs. SHAM; c, OVA + E2 vs. OVA.

Article Snippet: Antibody for adiponectin (26 kDa, 3553) was supplied by ProSci (Poway, CA, USA); antibodies for ADIPOR1 (43 kDa, sc-99183), ADIPOR2 (50 kDa, sc-46755), GAPDH (37 kDa, sc-365062), GLUT4 (53 kDa, sc-7938) and PPARG (53 kDa, sc-7196) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA); antibodies for AKT (60 kDa, cs-2920), pAKT (60 kDa, cs-4060) and APPL1 (82 kDa, cs-3858) were supplied by Cell Signaling (Danvers, MA, USA).

Techniques: Expressing

Journal: Endocrinology

Article Title: Growth hormone is necessary for the p53-mediated, obesity-induced insulin resistance in male C57BL/6J x CBA mice.

doi: 10.1210/en.2013-1220

Figure Lengend Snippet: Figure 4. Mechanisms underlying the increased expression of adipose p53. Quantitative PCR (mean values of 4 replicates) was used for evaluating the transcriptional regulation of adipose p53; p53 transcripts increased mainly in obese mice and, although to lesser extent, in Acro mice than in Lean animals (A). Whole extracts from adipose tissue of Obe, Lean, or Acro mice were used for Western blotting (B–D) or coimmunoprecipitation (E). The expression of total and phosphorylated p38 (B), that of Akt (C), and that of p44/42 (D) was evaluated by Western blot. For coimmunoprecipitation experiments, protein extracts were first immunoprecipitated with a p53 antibody, and after dissociation of immunocomplexes and protein resolution on SDS-PAGE, the level of expression of MDM2 protein associated with p53 was evaluated by a specific MDM2 antibody by Western blot (E). *, P .05; **, P .005, post hoc Tukey test. A.U., arbitrary units.

Article Snippet: The following antibodies were used: rabbit polyclonal antip53 (sc-6243), anti-p44/42 (sc-94), anti-GLUT4 (sc-7938), anti-pAKT1 (sc-7985-R), anti-sterol regulatory binding element (SREBP)-1 (sc-366), anti-suppressors of cytokine signal (SOCS)-3 (sc-9023), anti-signal transducers and activators of transcription (STAT)-1 (sc-464), anti-Tp-53-induced glycolysis and apoptosis regulator (TIGAR; sc-67273), anti-p21 (sc-397); goat polyclonal antihexokinase II (sc-6521); mouse monoclonal antiactin (sc-8432), anti-AKT1 (sc-5298), antimouse double-minute 2 homolog (MDM2; sc-965), anti-p38 (sc-7972), antiphosphop38 (sc-7973), anti-STAT5b (sc-1656), and anti-STAT3 (sc-8019) were obtained from Santa Cruz Biotechnology.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunoprecipitation, SDS Page

Journal: Endocrinology

Article Title: Growth hormone is necessary for the p53-mediated, obesity-induced insulin resistance in male C57BL/6J x CBA mice.

doi: 10.1210/en.2013-1220

Figure Lengend Snippet: Figure 6. Proposed mechanisms of GH in the regulation of insulin resistance. In conditions of inflammation of adipose tissue, for example, obesity (A), inflammatory cytokines, including IL-6, increase the expression of adipose SOCS1-STAT1- and SOCS3-STAT3-coupled proteins, leading to a higher sensitivity to normal circulating GH concentrations, which, in turns, increase the expression of the SOCS2-STAT5b system. The high intracellular levels of SOCS proteins allow an increased GHR signal through the p38 (and PI3k-Akt in Acro mice) pathways toward p53, increasing its expression and activity. Higher adipose levels of p53 trigger insulin resistance, affecting glucose and lipid metabolism, through regulation of key steps. Blocking GH receptor with a specific inhibitor (B) reduces the abnormal GH action in adipose tissue, ameliorating insulin resistance.

Article Snippet: The following antibodies were used: rabbit polyclonal antip53 (sc-6243), anti-p44/42 (sc-94), anti-GLUT4 (sc-7938), anti-pAKT1 (sc-7985-R), anti-sterol regulatory binding element (SREBP)-1 (sc-366), anti-suppressors of cytokine signal (SOCS)-3 (sc-9023), anti-signal transducers and activators of transcription (STAT)-1 (sc-464), anti-Tp-53-induced glycolysis and apoptosis regulator (TIGAR; sc-67273), anti-p21 (sc-397); goat polyclonal antihexokinase II (sc-6521); mouse monoclonal antiactin (sc-8432), anti-AKT1 (sc-5298), antimouse double-minute 2 homolog (MDM2; sc-965), anti-p38 (sc-7972), antiphosphop38 (sc-7973), anti-STAT5b (sc-1656), and anti-STAT3 (sc-8019) were obtained from Santa Cruz Biotechnology.

Techniques: Expressing, Activity Assay, Blocking Assay