Journal: PLoS ONE

Article Title: Arg-Gly-Asp (RGD)-Modified E1A/E1B Double Mutant Adenovirus Enhances Antitumor Activity in Prostate Cancer Cells In Vitro and in Mice

doi: 10.1371/journal.pone.0147173

Figure Lengend Snippet: Cells were grown and immunostained with CAR, integrin α v β 3 , or α v β 5 antibody and then subjected to flow cytometric analysis. Columns, percentages of cells expressing CAR, αvβ3 and αvβ5. Data are presented as means ± standard deviation for three independent experiments.

Article Snippet: To detect expression of CAR and integrin α v β 3 and α v β 5 , the cells were detached using an enzyme-free dissociation solution and then incubated with a mouse monoclonal anti-human CAR antibody (Upstate Biotechnology, Lake Placid, NY, USA), a mouse monoclonal anti-human integrin α v β 3 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), or a mouse monoclonal anti-human integrin α v β 5 antibody (R&D Systems, Inc., Minneapolis, MN, USA), all at a dilution of 1:100 for 1 h on ice.

Techniques: Expressing, Standard Deviation

Journal: Bone Research

Article Title: Ciliary parathyroid hormone signaling activates transforming growth factor-β to maintain intervertebral disc homeostasis during aging

doi: 10.1038/s41413-018-0022-y

Figure Lengend Snippet: PTH induces integrin α v β 6 expression to activate latent TGF-β. a Immunostaining images showing various types of integrin expressions in IVD tissue from 18-month-ld mice injected with PTH or vehicle and quantitative analysis ( b ). Scale bar, 50 μm. c qRT-PCR analysis of the mRNA levels of various integrin in NP tissue from 18-month-old mice injected with PTH or vehicle. Results reported as fold change. d Western blot analysis of integrin β 6 expression in NP cells of 18-month-old mice at different time points post PTH injection (PTH1-34, 100 nmol·L -1 ). e , f Chromatin immunoprecipitation assay with four different potential pCREB binding sites (primers 1, 2, 3 and 4) in the β6 integrin promoter. g pCREB, Integrin α V β 6 , pSmad2/3, or Safranin-O staining of IVD sections from an IVD ex vivo compression model of 30-month-old rat with treatment of either vehicle or PTH (PTH1-34, 100 nmol·L -1 ). Scale bar, 20 μm. h Quantitative analysis of the percentage of pCREB, pSmad2/3 positive cells and the Integrin α V β 6 positive areas as a percentage of total IVD area (Ar) of ( g ). All data are reported as the mean ± s.d. * P < 0.05. n = 8 per group. Statistical significance was determined by one-way ANOVA and Student's t -test

Article Snippet: Sections for immunostaining were processed using a standard protocol and incubated with primary antibodies to rabbit ACAN (Abcam, 1:100), CCN2 (Abcam, 1:100), integrin β 8 (Abcam, 1:200), pCREB (Abcam, 1:100), and PTH1R PRB-635P (Covance, 1:100), mouse pSmad2/3 (Santa Cruz, 1:100), integrin α v β 6 (Millipore, 1:100), integrin α v β 3 (Bioss, 1:100), integrin α v β 5 (Bioss, 1:100) at 4 °C overnight.

Techniques: Expressing, Immunostaining, Injection, Quantitative RT-PCR, Western Blot, Chromatin Immunoprecipitation, Binding Assay, Staining, Ex Vivo

Journal: EJNMMI Radiopharmacy and Chemistry

Article Title: Preclinical validations of [ 18 F]FPyPEGCBT- c (RGDfK): a 18 F-labelled RGD peptide prepared by ligation of 2-cyanobenzothiazole and 1,2-aminothiol to image angiogenesis

doi: 10.1186/s41181-016-0019-z

Figure Lengend Snippet: Uptake of [ 18 F]FPyPEGCBT- c (RGDfK) in cancer cells. U-87 MG and SKOV-3 cells were incubated with 100 kBq/mL [ 18 F]FPyPEGCBT- c (RGDfK) for 15 to 120 min at 37 or 4 °C ( a ). Cells were also co-treated with 10 −4 M c (RGDfV) for 1 h ( b ). Uptake of [ 18 F]FPyPEGCBT- c (RGDfK) was also evaluated after 1 h of incubation with cells with selective siRNA-mediated knocked down of α V , β 1 , β 3 or β 5 integrin subunits and indicated combinations ( c ). Depletions were verified by immunoblotting using specific antibodies as compared to untransfected cells (Unt) and to an irrelevant depletion of luciferase (si Luc). After incubation, cells were lysed and the cell-associated radioactivity was measured in a γ-counter. Data are means ± SD of at least three independent experiments performed in quadruplicate well. Statistical differences were analysed with the one-way ANOVA test followed by Dunnett’s post hoc test as compared to 37 °C controls ( b ) or to si Luc ( c ); # p < 0.001

Article Snippet: Promising peptide ligands with alternative binding motifs have recently been reported, including a) α V β 3 but not α V β 5 integrin (Zannetti et al. ); b) exclusively α 5 β 1 (Neubauer et al. ; Haubner et al. ); and c) exclusively α V β 6 (Hausner et al. ).

Techniques: Incubation, Western Blot, Luciferase, Radioactivity