Journal:

Article Title: Transient alteration of T cell fine specificity by a strong primary stimulus correlates with T cell receptor down-regulation

doi:

Figure Lengend Snippet: Inhibition of protein tyrosine kinase activity blocks TCR down-regulation. P14 TCR transgenic spleen cells (A, B) or P14.G5 CTL clone (C, D) were preincubated for 60 min in the presence of PP1 or genistein. Preincubation with herbimycin A was done overnight with P14 spleen cells or for 60 min with P14.G5 CTL. EL-4 cells pulsed with p33 (1 μM) were then added for 4 h and cells stained for flow cytometry with anti-CD8 and anti-Vα2 antibodies (A, C) or with anti-CD8 and anti-CD69 antibodies (B, D). Mean fluorescence intensities are expressed as % or MFI of control in the absence of peptide.

Article Snippet: P14.G5 CTL (6×10 4 /well) or P14 spleen cells (5×10 5 /well) were preincubated for 60 min with the src family kinase-specific inhibitor PP1 (10 μM and 50 μM, respectively) (Bio-mol, Plymouth Meeting, PA) [ 28 ].

Techniques: Inhibition, Activity Assay, Transgenic Assay, Staining, Flow Cytometry, Fluorescence, Control

Journal: The Journal of Biological Chemistry

Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation *

doi: 10.1074/jbc.M113.515254

Figure Lengend Snippet: Activation of IGF-1R in α6 integrin signaling complexes is dependent on the kinase activity of Src family kinases. Primary lens cells in culture were exposed to the highly specific Src family kinase inhibitor, PP1, or to the vehicle dimethyl sulfoxide (DMSO) for 4 h. Following PP1 treatment, cells were immunoprecipitated for α6 integrin and the immunoprecipitates were immunoblotted for α6 integrin and phospho-Src (p-Src) (A), Src (B), phospho-IGF-1R (p-IGF-1R) (C), and IGF-1R (D). A–D, graphical representation of densitometric analysis of co-immunoprecipitation studies showing the ratio of activated, p-Src/α6 integrin (A), total Src/α6 integrin (B), activated p-IGF-1R/α6 integrin (C), and total IGF-1R/α6 integrin (D). Data shows that blocking activation of Src kinases prevented α6 integrin transactivation of IGF-1R. Results shown are quantified from more than three independent studies. Error bars represent S.E. *, p < 0.05, t test.

Article Snippet: For blocking activation of Src family kinases, cells were exposed to the Src family kinase-specific inhibitor PP1 (10 μ m , Enzo Life Sciences, Farmingdale, NY) for 4 h. Controls were treated with the vehicle dimethyl sulfoxide.

Techniques: Activation Assay, Activity Assay, Immunoprecipitation, Blocking Assay