Journal: Molecular neurobiology

Article Title: Synaptic ERK2 Phosphorylates and Regulates Metabotropic Glutamate Receptor 1 In Vitro and in Neurons

doi: 10.1007/s12035-016-0225-4

Figure Lengend Snippet: ERK-mediated phosphorylation of mGluR1a in rat cerebellar neurons. a–c Phosphorylation of mGluR1a at ERK-preferred motifs, including S/TP (a), TP (b), and PXSP (c). Cerebellar mGluR1a was immunopurified by the anti-mGluR1a antibody. Phosphorylation signals at three phosphomotifs (pS/TP, pTP, and PXpSP) in immunopurified mGluR1a were detected by immunoblots (IB) with indicated antibodies. d Dephosphorylation of mGluR1a PXSP phosphorylation by λ-protein phosphatase (λ-PP). Cerebellar slices were incubated with λ-PP (200–400 units) for 1 h at 30 °C. Note that PXSP phosphorylation was dephosphorylated by λ-PP. e Representative immunoblots illustrating effects of U0126 on phosphorylation of mGluR1a at PXSP and TP motifs and on pERK1/2-mGluR1a and ERK1/2-mGluR1a interactions in cerebellar neurons. f, g Quantifications of effects of U0126 on mGluR1a PXSP (f) and TP (g) phosphorylation. h Quantifications of effects of U0126 on pERK2-mGluR1a and ERK2-mGluR1a interactions. Cerebellar slices were incubated with U0126 (0.5 or 5 µM) for 30 min at 30 °C (e–h). Slices were collected for mGluR1a immunoprecipitation (IP). Phosphorylation levels of mGluR1a at PXSP and TP and proteins bound to immunopurified mGluR1a (pERK1/2 and ERK1/2) were visualized by immunoblots. Values in graphs were measured as ratios of PXpSP to mGluR1a (f), pTP to mGluR1a (g), pERK2 to mGluR1a (h), and ERK2 to mGluR1a (h). All values were analyzed by one-way ANOVA: PXpSP (f), F(2, 7) = 5.05, p < 0.05; pTP (g), F(2, 7) = 0.26, p > 0.05; pERK2 (h), F(2, 7) = 37.44, p < 0.05; and ERK2 (h), F(2, 7) = 14.67, p < 0.05. Data are presented as means ± SEM (n = 3–4 per group). *p < 0.05 versus vehicle

Article Snippet: The beads were then divided into two tubes, and λ-protein phosphatase (200–400 units, New England Biolabs), a serine/threonine/tyrosine dual phosphatase, was added to one of tubes.

Techniques: Western Blot, De-Phosphorylation Assay, Incubation, Immunoprecipitation

Journal:

Article Title: Composite Low Grade B-Cell Lymphomas with Two Immunophenotypically Distinct Cell Populations Are True Biclonal Lymphomas

doi:

Figure Lengend Snippet: Antibodies Used in this Study

Article Snippet: The rest of the staining procedure was performed on the Ventana immunostainer. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 CD/name (clone) Reactivity Source Dilution CD3 polyclonal T cells Dako 1:100 CD5 (4C7) T cells, B-sub Novocastra 1:50 CD10 (56C6) cALLA antigen Novocastra 1:40 CD20 (L26) B cells Dako 1:200 CD21 (1F8) FDC, B-sub Dako 1:20 CD23 (Bu38) B-sub, FDC Binding Site 1:200 CD43 (Leu22) T cells, B-sub BD 1:50 IgD polyclonal IgD Dako 1:400 kappa polycl. κ light chains Dako 1:25,000 lambda polycl. λ light chains Dako 1:25,000 MIB1 Ki67 antigen Immunotech 1:40 bcl-2 (124) bcl-2 protein Dako 1:20 cyclin D1 (P2D11F11) cyclin D1 protein Novocastra 1:10 p53 (DO7) p53 protein Dako 1:50 p27 (Kip-1) p27/kip1 protein Transduction 1:1000 Open in a separate window BD, Becton Dickinson, Mountain View, CA; Binding Site, The Binding Site, Birmingham, UK; Dako, Dako Corp., Carpinteria, CA; Immunotech, Immunotech Inc., Westbrook, ME; Novocastra, Novocastra, Newcastle, UK; Transduction, Transduction Laboratories, Lexington, KY. Antibodies Used in this Study

Techniques: Binding Assay, Transduction