dok 1  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: The lentiviral vectors containing DOK-1 specific ShRNA (DOK_ShRNA) or DOK-1 cDNA (DOK) were administrated together with controls (empty vector or vector containing non-specific scrambled ShRNA) before OVA challenge. (A) Representative western blotting demonstrating DOK-1 protein expression in the lungs of the mice. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression (B) Immunofluorescent staining on the tissue sections from the lungs using Alexa Fluor 488-conjugated DOK-1 antibody and DAPI stains.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: shRNA, Plasmid Preparation, Western Blot, Expressing, Over Expression, Staining

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: (A) Lung sections were stained with hematoxylin and eosin, D-PAS, alcian blue for the evaluation of inflammatory cells and airway mucus responses. ×40 of original magnification. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression. At least 4 mice were included in each group. (B) Inflammatory index that scored parenchymal inflammation. At least 4 mice were included in each group. *P<0.05 (C) Mucus index evaluated by morphometric analysis representing alcian blue stained mucus cells (percentage of positive cells) in airway epithelial cells. At least 4 mice were included in each group. *P<0.05.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: Staining, Plasmid Preparation, shRNA, Over Expression

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: (A) FACS Histogram analysis on BAL cells from the mice after OVA sensitization and challenge. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression. Each lane indicates CD4 + T cell population stained with Cy5-anti-CD4 antibody. (B) BAL CD4(+)T cells gated with PE-conjugated CD4 were further evaluated by intracellular staining against Cy5-conjugated-IL-4 and FITC-conjugated-IFN-γ. (C) The levels of inflammatory Th1 and Th2 cytokines and a chemokine in BAL fluid were measured by ELISA at 24 hrs after the last OVA challenge. Data represent means ± S.E.M. Each group contains at least 7 mice. *P<0.05, ***P<0.001 vs. OVA-challenged mice.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: Plasmid Preparation, shRNA, Over Expression, Staining, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: Expression and/or activation (phosphorylation) of STAT6, STAT4, GATA3, and T-bet transcriptional factors were evaluated by Western blot analysis. A representative gel photo out of 6 similar independent experiments. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: Expressing, Activation Assay, Western Blot, Plasmid Preparation, shRNA, Over Expression

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: A representative gel photo out of five similar independent experiments. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: Plasmid Preparation, shRNA, Over Expression

Journal: PLoS ONE

Article Title: An Essential Regulatory Role of Downstream of Kinase-1 in the Ovalbumin-Induced Murine Model of Asthma

doi: 10.1371/journal.pone.0034554

Figure Lengend Snippet: (A) Lung sections were stained with Alexa Fluor 488-conjugated DOK-1 and Alexa Fluor 568-conjugated STAT-6 antibodies and DAPI stain. (B) Lung sections were stained with Alexa Fluor 488-conjugated DOK-1 and Alexa Fluor 568-conjugated STAT-4 antibodies and DAPI stains. A representative photo of 7 similar experiments. Con, non OVA challenged; OVA, OVA-challenged with empty lentiviral vector; DOK_ShRNA, OVA-challenged with DOK_ShRNA knockdown; DOK, OVA-challenged with DOK-1 overexpression.

Article Snippet: DOK-1 cDNA was cloned by PCR amplification using the following primers : 5′-CTACCTGAGCTACCAGTCCGC-3′ (Sense) and 5′-CGTGAAGAATGTGCGAGAC-3′ (antisense) (Genecopoeia, Germantown, MD).

Techniques: Staining, Plasmid Preparation, shRNA, Over Expression

Journal: Frontiers in Immunology

Article Title: Complement Receptor 3-Mediated Inhibition of Inflammasome Priming by Ras GTPase-Activating Protein During Francisella tularensis Phagocytosis by Human Mononuclear Phagocytes

doi: 10.3389/fimmu.2018.00561

Figure Lengend Snippet: Dok-1 is phosphorylated during infection with serum-opsonized Schu S4 and co-localizes with Ras GTPase-activating protein (RasGAP). Monocyte-derived macrophage monolayers were infected with non-opsonized or pre-opsonized bacteria (MOI of 100). Infection was synchronized at 4°C followed by incubation at 37°C. At the indicated time points, cell lysates were collected and subjected to Western blot for phosphorylated Lyn and Dok-1 (A) . Phosphorylated Dok-1/actin band intensity ratio at different time points from (A) is shown in (D) . Association of RasGAP with phosphorylated Dok-1 was examined by immunoprecipitation using anti-RasGAP Ab (B) . Phosphorylated Dok-1/RasGAP band intensity ratio at different time points from (B) is shown in (D) . Data from (A) and (B) are representative of 3 independent experiments. Co-localization of RasGAP with Dok-1 was examined at the 5 min time point by confocal microscopy (C) . Data are representative photomicrographs from three independent experiments.

Article Snippet: Phospho-Dok-1 (pTyr362) was from Acris Antibodies, Inc. (San Diego, CA, USA).

Techniques: Infection, Derivative Assay, Incubation, Western Blot, Immunoprecipitation, Confocal Microscopy