Journal: Oxidative Medicine and Cellular Longevity

Article Title: Prevention of Streptozotocin-Induced Diabetic Nephropathy by MG132: Possible Roles of Nrf2 and I κ B

doi: 10.1155/2017/3671751

Figure Lengend Snippet: Effects of MG132 on diabetes-induced renal fibrosis in wild-type mice (a) and Nrf2-KO mice (b) were determined by detecting the expression of FN and TGF- β with western blotting assay. Diabetes-induced fibrotic changes (fold) between wild-type and Nrf2-KO mice and the decreased percentages of these changes with MG132 between WT and Nrf2-KO diabetic mice were compared (c). Data are presented as mean ± SD. ∗ p < 0.05 versus WT/control or Nrf2-KO/control correspondingly; # p < 0.05 versus WT/DM or Nrf2-KO/DM correspondingly; & p < 0.05 versus WT mice.

Article Snippet: The primary antibodies were FN (1 : 200 dilution), TGF- β (1 : 1000 dilution), 3-NT (1 : 1000 dilution), 4-HNE (1 : 1000 dilution), IL-6 (1 : 500 dilution), NF- κ B (1 : 1000 dilution), I κ B- α (1 : 1000 dilution), Nrf2 (1 : 500 dilution), actin (1 : 3000 dilution), and α -tubulin (1 : 2000 dilution), all of which were purchased from Santa Cruz Biotechnology except for 3-NT (Millipore), 4-HNE (Alpha Diagnostic), and TGF- β , NF- κ B, I κ B- α , and α -tubulin (Cell Signaling).

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: A critical role of platelet TGF-β release in podoplanin-mediated tumour invasion and metastasis

doi: 10.1038/srep42186

Figure Lengend Snippet: ( a ) Measurement of TGF-β1 or PDGF-BB concentrations in tumour-platelet reactants using Bio-Plex suspension array system. Error bars indicate standard deviation (SD). **P < 0.01 by Student’s t test. ( b , c ) Morphological and physiological changes in cells after treatment with or without 3 ng/ml recombinant TGF-β1 for 48 h. ( b ) Cells were stained for E-cadherin (green), F-actin (red; phalloidin) and nuclear DNA (blue; Hoechst 33342). Scale bars represent 50 μm. ( c ) Cell lysates were immunoblotted with antibodies to N-cadherin, claudin-1, podoplanin (PDPN, clone D2-40) and TopoIIβ. ( d ) Cells were either left untreated or treated with supernatants of platelets alone (platelets), supernatants of platelet–cell reactants (platelets + cells), or 3 ng/ml of recombinant TGF-β1 for 0.5 h. The cell lysates were immunoblotted with antibodies against phospho-Smad2/3 (pSmad2/3), Smad3, and TopoIIβ.

Article Snippet: Proteins were then transferred to Immobilon-P polyvinylidene fluoride (PVDF) membranes (Merck Millipore, Darmstadst, Germany) and immunoblotted with antibodies against human podoplanin (D2–40; AbD Serotec, Kidlington, UK or Dako, Glostrup, Denmark), N-cadherin (Cell Signaling Technology, Danvers, MA, USA), Claudin-1 (Cell Signaling Technology), pSmad3 (Cell Signaling Technology), pSmad2/3 (Cell Signaling Technology), Smad3 (Cell Signaling Technology), TopoIIβ (clone 40; BD Transduction Laboratories, Washington, DC, USA) and β-actin (clone AC-15; Santa Cruz, Dallas, TX, USA).

Techniques: Standard Deviation, Recombinant, Staining

Journal: Oncology Reports

Article Title: Downregulation of miR-505-3p predicts poor bone metastasis-free survival in prostate cancer

doi: 10.3892/or.2018.6826

Figure Lengend Snippet: miR-505-3p inhibits the activation of TGF-β signaling by targeting SMAD2 and SMAD3 in PCa cells. (A-C) GSEA revealed that miR-505-3p downregulation was positively associated with TGF-β signaling activity. (D) Transcriptional activity based on a TGF-β/Smad-responsive luciferase reporter as assessed in the indicated cells. *P<0.05. (E) Western blotting of nuclear pSMAD2/3 expression. The nuclear protein p84 was used as a nuclear protein marker. (F-H) Real-time PCR analysis of downstream target gene expression of TGF-β signaling, including IL11, PTHRP and CTGF in the indicated cells. *P<0.05. PCa, prostate cancer; GSEA, gene set enrichment analysis.

Article Snippet: Antibodies against SMAD2 (cat. no. 5339), SMAD3 (cat. no. 9523), pSMAD2/3 (cat. no. 9510) and SMAD2/3 (cat. no. 8685) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), and p84 (cat. no. PA5-27816) was obtained from Invitrogen; Thermo Fisher Scientific, Inc. All antibodies aforementioned were diluted 1:1,000.

Techniques: Activation Assay, Activity Assay, Luciferase, Western Blot, Expressing, Marker, Real-time Polymerase Chain Reaction