Journal: Science Advances

Article Title: Cardiomyocyte-derived YOD1 promotes pathological cardiac hypertrophy by deubiquitinating and stabilizing STAT3

doi: 10.1126/sciadv.adu8422

Figure Lengend Snippet: To verify the effect of YOD1 on STAT3, the STAT3 inhibitor Stattic (10 mg/kg, gavage) or vehicle [0.5% O -carboxymethylcellulose (CMC)–Na and 0.25% Tween 80] was administered every 3 days via oral gavage. Healthy male YOD1CKO mice aged 6 to 8 weeks and YOD1 fl/fl mice were injected with Ang II (1 μg kg –1 min –1 ) or normal saline via an osmotic pump (catalog no. ALZET Model 1004, USA) for 4 weeks to induce cardiac hypertrophy. ( A ) Representative M-mode echocardiography of mice in each group. ( B and C ) Myocardial function parameters, including EF (B) and FS (C), were evaluated in mice through echocardiography. ( D ) The plasma levels of ANP in each group. ( E ) Representative images of whole hearts. Scale bars, 2.5 mm. ( F ) Representative images of H&E staining of transverse section in myocardium tissues. Scale bars, 2.5 mm and 50 μm. ( G and H ) Representative images of WGA (G) staining in sections of hearts and quantitative area analysis (H). Scale bars, 50 μm. ( I to L ) Fibrotic areas were evaluated using Masson’s trichrome (I) and Sirius Red (K) staining, as well as the quantification of fibrotic regions (H and J) in heart sections from each group. Scale bars, 50 μm. ( M and N ) Representative Western blot analysis of MyHC and ANP (M) and densitometric quantification (N). n = 6. n.s., P > 0.05; *** P < 0.001.

Article Snippet: Ang II (catalog no. HY-13948), Stattic (catalog no. HY-13818), and CHX (catalog no. HY-12320) were obtained from MedChemExpress (New Jersey, USA).

Techniques: Injection, Saline, Clinical Proteomics, Staining, Western Blot

Journal: Journal of neurochemistry

Article Title: Phosphatidylinositol-3 kinase mediates the sweet suppressive effect of leptin in mouse taste cells

doi: 10.1111/jnc.15268

Figure Lengend Snippet: Effect of bath applied leptin on sucrose responses of taste receptor family 1 member 3 (T1R3)-green fluorescent protein (GFP) taste cells (TCs) from mouse fungiform papillae. (a) Sample recordings of TC responses to 500 mM sucrose (Suc) before (cont), during (leptin) and after (wash) treatment with 20 ng/ml leptin. The picture shows a T1R3-GFP TC from which taste responses were recorded. (b). Responses of 10 individual T1R3-GFP TCs to 500 mM sucrose before (left), during (middle) and after (right) treatment of 20 ng/ml leptin. (c) Effects of leptin on summated responses of T1R3-GFP TCs to 500 mM sucrose (n = 10 cells/ 9 mice, F = 14.869, p < .001, repeated ANOVA). Values are means ± SE. (d–i) Phosphoinositide 3-kinase (PI3K) inhibitors suppressed leptin’s suppression of TC responses to sucrose. (d, g) Sample recordings of TC responses to 500 mM sucrose (Suc) before (cont), during (leptin) and after (wash) treatment with 20 ng/ml leptin. PI3K inhibitor wortmannin (30 nM, d) or LY294002 (30 μM, g) was added to bath solution throughout the experiment. The picture shows a T1R3-GFP TC from which taste responses were recorded. (e, h) Responses of 8 and 7 individual T1R3-GFP TCs to 500 mM sucrose before (left), during (middle) and after (right) treatment of 20 ng/ml leptin under the existence of 30 nM wortmannin (e) or 30 μM LY294002 (h). (f, i) The effects of leptin on summated responses of T1R3-GFP TCs to 500 mM sucrose under the existence of 30 nM wortmannin (n = 8 cells/ 7 mice, F = 0.3577, p > .1, repeated ANOVA, f) or 30 μM LY294002 (n = 7 cells/ 7 mice, F = 0.0375, p > .1, repeated ANOVA, i). Values are means ± SE. (j–o) Signal transducer and activator of transcription 3 (STAT3) inhibitor and Src homology region 2 domain-containing phosphatase-2 (SHP2) inhibitor did not affect leptin’s suppression of TC responses to sucrose. (j, m) Sample recordings of TC responses to 500 mM sucrose (Suc) before (cont), during (leptin) and after (wash) treatment with 20 ng/ml leptin. STAT3 inhibitor stattic (10 μM, i) or SHP2 inhibitor SHP099 (10 μM, m) was added to bath solution throughout the experiment. The picture shows a T1R3-GFP TC from which taste responses were recorded. (k, n) Responses of 8 individual T1R3-GFP TCs to 500 mM sucrose before (left), during (middle) and after (right) treatment of 20 ng/ml leptin under the existence of 10 μM stattic (k) or 10 μM SHP099 (n). (l, o) Effects of leptin on summated responses of T1R3-GFP TCs to 500 mM sucrose under the existence of 10 μM stattic (n = 8 cells/ 8 mice, F = 4.0694, p < .05, repeated ANOVA, l) or 10 μM SHP099 (n = 8 cells/ 7 mice, F = 5.8569, p < .05, repeated ANOVA, o). Values are means ± SE. **p < .01, *p < .05, post hoc paired t-test with Holm correction

Article Snippet: Recombinant murine leptin (20 ng/ml; PeproTech, Cat# 450-31), wortmannin (10 nM; Tocris Bioscience, Cat# 1,232), LY294002 (30 μM; Sigma-Aldrich, Cat# L9908), SHP099 (10 μM; Cayman Chemical, Cat# 20,000), and stattic (10 μM; Axon Medchem, Cat# 2,314) were applied from the basolateral side of TCs.

Techniques:

Journal: International Journal of Oncology

Article Title: STAT3 activation in HER2-overexpressing breast cancer promotes epithelial-mesenchymal transition and cancer stem cell traits

doi: 10.3892/ijo.2013.2195

Figure Lengend Snippet: STAT3 inhibitor, Stattic abolishes the stem cell marker expression. (A) Dose response curve of MCF7-HER2 for the STAT3 inhibitor Stattic. (B) Stattic treatment abolished pSTAT3 and the stem cell marker expression in MCF7-HER2. The EMT driver slug expression was also abolished upon Stattic treatment. (C) Western blot analyses showed that, with Stattic treatment, expression of vimentin and slug was downregulated while E-cadherin expression was upregulated in MCF7-HER2. (D) Combined treatment of Herceptin and Stattic showed a synergistic cell growth inhibition effect on HER2-overexpressing, ER-positive breast cancer cells.

Article Snippet: Cells were allowed to grow until about 50% confluence, and then treated with the following dosage treatments: untreated control, Stattic (5 μ M), Herceptin (10 μ g/ml, a gift of Genentech) and Stattic (5 μ M) + Herceptin (10 μ g/ml) combination.

Techniques: Marker, Expressing, Western Blot, Inhibition