Journal: Journal of Clinical Medicine

Article Title: Current Concepts on the Reno-Protective Effects of Phosphodiesterase 5 Inhibitors in Acute Kidney Injury: Systematic Search and Review

doi: 10.3390/jcm9051284

Figure Lengend Snippet: Animal studies evaluating the potential reno-protective effects of vardenafil.

Article Snippet: [ ]/ Germany/2008 , Sprague Dawley rats , Mouse monoclonal anti-Thy 1 antibody ER-4 Single injection, 1 mg/kg , Vardenafil Orally 20 mg/kg within 18 h and 10 mg/kg/day for 48 h , PRE and POST , 24-h urine collection on Days 2 and 6 Blood sample: Day 6 , ↑PDE5-A, ⇔sCr ↑proteinuria Mesangial proliferation , ↑cGMP, ↓TSP-1, ⇔sCr ↓proliferation/cell number(glomerular) ↓collagen IV/fibronectin (glomerular) ↓TGF-β activation ⇔proteinuria , POS.

Techniques: Animal Model, Injection, Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Mechano-transduction in Osteoblastic Cells Involves Strain-regulated Estrogen Receptor α-mediated Control of Insulin-like Growth Factor (IGF) I Receptor Sensitivity to Ambient IGF, Leading to Phosphatidylinositol 3-Kinase/AKT-dependent Wnt/LRP5 Receptor-independent Activation of β-Catenin Signaling *

doi: 10.1074/jbc.M109.027086

Figure Lengend Snippet: ERα sensitizes UMR-106 osteoblasts to IGF-I in an estrogen-independent fashion requiring Src activity and physically associates with IGF-IR. a , whole cell lysates prepared from UMR-106 cells pretreated with vehicle (ethanol) or ICI 182,780 for 16 h prior to being treated with 1, 0.1, 0.5, 1, 5, or 10 ng/ml IGF1 for 3 h. The levels of total and phospho-AKT and GSK3-β as well as total and active β-catenin were analyzed by Western blotting. b , scanning densitometry was performed on Western blots from three independent experiments, and the results are represented as fraction of the control. Values shown are mean ± S.E. ***, p < 0.001; **, p < 0.01; *, p < 0.05 by one-way ANOVA with Bonferroni post hoc analysis. p values are only shown in the des-(1–3)IGF-I group when they from the 10 ng/ml des-(1–3)IGF-I + ICI 182,780. c , whole cell lysates prepared from UMR-106 cells were pretreated with vehicle ( Veh. ) (ethanol) or ICI 182,780 for 16 h, E 2 (3 h), or the pan-Src inhibitor PP2 (30 min) prior to being treated with 1 ng/ml des-(1–3)IGF1 for 1 h. The levels of total and phospho-AKT were measured by Western blotting. d , nondenaturing TLB lysates were prepared from UMR-106 cells treated with 50 ng/ml IGF1 for 3 h. 1 mg of extract was immunoprecipitated with 5 μg of mouse IgG or SR1000 mouse monoclonal antibody recognizing ERα. Immunoprecipitates were analyzed by Western blotting to determine the levels of ERα in the immunoprecipitates and the amount of IGF-IR associated with ERα. Also shown are Western blots for 10 μg of “input” lysate to determine the base-line expression of both ERα and IGF-IR in the lysates.

Article Snippet: The mouse monoclonal antibody recognizing ERα (SRA1000) was purchased from Cambridge Bioscience (Cambridge, UK).

Techniques: Activity Assay, Western Blot, Control, Immunoprecipitation, Expressing

Journal: American Journal of Translational Research

Article Title: 4-Hydroxy estrogen metabolite, causing genomic instability by attenuating the function of spindle-assembly checkpoint, can serve as a biomarker for breast cancer

doi:

Figure Lengend Snippet: A. Immunohistochemical analysis of four groups mice. H&E staining of breast in mice from C57BL/6J group, C57BL/6J + E2 group, C57BL/6J-CYP1B1 group and C57BL/6J-CYP1B1 + E2 group. 400×, bar = 50 μm. B. Representative images for H&E staining of breast, liver and lung samples from four groups. B. Expression of ki67 and ER protein in mammary gland of four groups mice. Cells stained brown indicate positive expression. Mammary gland from C57BL/6J group, C57BL/6J + E2 group, C57BL/6J-CYP1B1 group and C57BL/6J-CYP1B1 + E2 group mice were stained for Ki67 and ERα.

Article Snippet: The sections were incubated with specific mouse monoclonal antibodies against ER, PR, HER-2 and Ki67 (American Diagnostica Inc.) at 25°C for 30 min, followed by incubation with the biotin-conjugated secondary anti-mouse antibodies (Dako, Glostrup, Denmark).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal:

Article Title: Attenuation of estrogen receptor ?-mediated transcription through estrogen-stimulated recruitment of a negative elongation factor

doi: 10.1101/gad.1214104

Figure Lengend Snippet: NELF in breast cancer cell lines and normal breast tissues. (A) Control, COBRA1, and NELF-E siRNA knockdown cells were grown in the Matrigel-containing three-dimensional tissue culture system in the presence or absence of E2 (10 nM). After 3 wk of growth incubation, the images were observed under phase microscopy with a 10× objective. (B) COBRA is preferentially expressed in the luminal epithelial cells of the mammary gland. Immunostaining of normal mammary ducts for COBRA1 (right) and ERα (center). The exclusive staining for smooth muscle actin (SMA) in myoepithelial cells is shown as a control (left). The brown color indicates the presence of protein. The locations of luminal epithelial (L) and myoepithelial cells (M) are also indicated.

Article Snippet: The following antibodies were used: a mouse monoclonal antibody against ERα (Novo Castra Laboratories) at 1:80 dilution, a mouse monoclonal antibody to SMA (BioGenex) at 1:1600 dilution, and rabbit polyclonal antisera against COBRA1, 1:50 dilution.

Techniques: Incubation, Microscopy, Combined Bisulfite Restriction Analysis Assay, Immunostaining, Staining