Journal: Antioxidants

Article Title: Overexpression of Mitochondrial Ferritin Enhances Blood–Brain Barrier Integrity following Ischemic Stroke in Mice by Maintaining Iron Homeostasis in Endothelial Cells

doi: 10.3390/antiox11071257

Figure Lengend Snippet: Overexpression of FtMt abrogates endothelial cell apoptosis after cerebral I/R. Western blot and densitometric analyses of ( A ) the ratio of Bcl-2 to Bax, ( B ) the ratio of cleaved caspase-3 to procaspase-3 ( n = 4). The data are expressed relative to the mean value in the WT Con group ( n = 4). The results are presented as the mean ± SEM. ** p < 0.01.

Article Snippet: The following primary antibodies were used: anti-β-actin (1:10,000) and anti-transferrin receptor 1 (TfR1; 1:2000) were obtained from Sigma-Aldrich (St. Louis, MO, USA) (#A5441, #SAB4200398); anti-ferroportin1 (FPN1; 1:5000) and anti-divalent metal transporter 1 (DMT1±IRE; 1:5000) were obtained from Alpha Diagnostic International (San Antonio, TX, USA) (#MTP11-S, #NRAMP21-S, #NRAMP23-S); anti-4-hydroxynonenal (4-HNE), anti-mitochondrial ferritin (1:5000), anti-ferritin light chain (FtL; 1:10,000) and anti-ferritin heavy chain (FtH; 1:10,000) were obtained from Abcam (#ab46545, #ab66111, #ab109373, #ab183781); anti-caspase3 (1:5000), anti-Bcl-2 (1:2000) and anti-Bax (1:2000) were obtained from Cell Signaling Technology (Danvers, MA, USA) (#14220, #3498, #2772); anti-ZO-1 (1:2000), anti-occludin (1:2000) and anti-claudin-5 (1:2000) were obtained from Thermo Fisher (Waltham, MA, USA) (#PA5-85256, #40-4700, #34-1600).

Techniques: Over Expression, Western Blot

Journal: The American Journal of Pathology

Article Title: A 2A Adenosine Receptor (A 2A AR) as a Therapeutic Target in Diabetic Retinopathy

doi: 10.1016/j.ajpath.2011.01.018

Figure Lengend Snippet: Role of A2A adenosine receptor (A2AAR)-cAMP signaling in the mitogen-activated protein (MAP) kinase pathway. (A) Effect of A2AAR activation on Amadori-glycated albumin (AGA)-induced extracellular signal-regulated kinase (ERK) MAP kinase activation. Retinal microglial cells were treated with AGA (500 μg/mL) in the presence or absence of CGS21680 (20 and 40 μmol/L) for 4 hours. Phospho-ERK (p-ERK) and total ERK MAP kinase in cell lysates were determined using Western blot analysis. Data shown are the mean + SD of three experiments. (B, C) Effect of A2AAR activation on AGA-induced C-Raf (B) and B-Raf (C) activation. Retinal microglial cells were treated with AGA (500 μg/mL) in the presence or absence of CGS21680 (20 μmol/L) or 8-pCPT-cAMP (250 μmol/L) for the indicated times (0 to 60 minutes). Phospho-C-Raf (Ser338), phospho-B-Raf, total C-Raf, and B-Raf in the cell lysates were determined using Western blot analysis.

Article Snippet: Antibodies for β-actin (Sigma), phospho-C-Raf (Ser338), phospho-B-Raf, C-Raf, B-Raf, phospho-ERK, and ERK mitogen-activated protein kinase (Cell Signaling Technology, Beverly, MA) were detected with a horseradish peroxidase-conjugated antibody and enhanced chemiluminescence (ECL) (Amersham BioSciences, Buckinghamshire, UK).

Techniques: Activation Assay, Western Blot

Journal: Molecular Cancer

Article Title: Targeting renal cell carcinoma with NVP-BEZ235, a dual PI3K/mTOR inhibitor, in combination with sorafenib

doi: 10.1186/1476-4598-10-90

Figure Lengend Snippet: NVP-BEZ235 and Sorafenib inhibit the growth of renal cancer cell lines . A, 786-0 and Caki-1 cellular growth was monitored with colorimetric MTS assay after 48 hours of treatment with NVP-BEZ235 (NVP, 1 μM) or Sorafenib (Soraf, 10 μM) either alone or in combination, or DMSO (Control). Columns, mean cell viability relative to control of four independent experiments; bars, SD. *, P < 0.05, **, P < 0.01, #, P < 0.001 compared to control, or otherwise as specified by brackets. B, NVP-BEZ235 inhibits PI3K/mTOR pathway in renal cancer cells. 786-0 and Caki-1 cells were treated with increasing doses of NVP-BEZ235 or DMSO (Control) for 4 hours. Cells were subsequently lysed and lysates were examined for phospho-Akt (Ser 473), Akt, phospho-S6 (Ser 235/236) or S6 expression level by Western blot analysis. C, 786-0 and Caki-1 cells were treated with increasing doses of sorafenib or DMSO as a control for 4 hours. Cells were processed as under panel B and analyzed for phospho-MAPK and MAPK (Thr202/Tyr204). The illustrated blots are representative of three similar experiments.

Article Snippet: Antibodies directed against phospho-Akt (S473), Akt, phospho-S6 ribosomal protein (Ser235/236), S6 ribosomal protein, phospho-MAPK (Thr202/Tyr204), MAPK, cleaved caspase-3 and actin were from Cell Signaling.

Techniques: MTS Assay, Expressing, Western Blot

Journal: Molecules (Basel, Switzerland)

Article Title: Extract from Black Soybean Cultivar A63 Extract Ameliorates Atopic Dermatitis-like Skin Inflammation in an Oxazolone-Induced Murine Model.

doi: 10.3390/molecules27092751

Figure Lengend Snippet: Figure 6. A63 extract prevents eosinophil infiltration toward the inflammatory site via CCL26 expression downregulation and the inactivation of JAK1/STAT6 signaling. qPCR analysis of CCL26 expression in IL-4/IL-13-induced HS68 cells (A). Western blot results of IL-4/IL-13-induced HS68 cells (B). Results are expressed as mean ± standard deviation (### p < 0.001 versus control group, *** p < 0.001 versus IL-4/13 treated group).

Article Snippet: The membranes were blocked with a 5% BSA solution and treated with primary antibodies for anti-phospho-p44/42 MAPK (ERK1/2), phospho-p38 MAPK, phospho-SAPK/JNK, phospho-STAT3, phospho-STAT6, phospho-JAK1, p44/42 MAPK (ERK1/2), p38 MAPK, JNK, STAT3, STAT6, JAK1, and GAPDH (Cell Signaling Technology, MA, USA, 1:1000).

Techniques: Expressing, Western Blot, Standard Deviation, Control