Journal: bioRxiv

Article Title: Reticular adhesions: A new class of adhesion complex that mediates cell-matrix attachment during mitosis

doi: 10.1101/234237

Figure Lengend Snippet: (A-M) Confocal images of U2OS cells plated for 3 h on vitronectin and immuno-labeled against integrin β5 and: the alpha V (aV) subunit of the αVβ5 heterodimer (A); consensus adhesome components [paxilin (B), FAK (C), zyxin (D), kindlin 2 (E)]; integrin β 5-binding partners [CSK (F), ICAP1 (G), DOK1 (H)]; phospho-tyrosine (I); and cytoskeletal proteins [F-actin (J), cytokeratin (K), beta (β)-tubulin (L), vimentin (M)]. (N) Confocal image of U2OS cells plated for 3 h on laminin (ECM ligand not bound by αVβ5) and immuno-labeled for integrin β5 and vinculin. (O) Confocal images of an unpermeabilised U2OS cell expressing β5-2GFP, immuno-labeled for β5. Scale bars: 10 μm. Boxed areas shown at higher magnification in lower right corners.

Article Snippet: Primary antibodies used for immunofluorescence and/or immuno-blotting include: anti-integrin β5 (15F11; MAB2019Z) (Millipore); anti-integrin αVβ5 (P1F6) (Abcam); polyclonal (rabbit) anti-integrin β5 (ab15459) (Abcam); anti-integrin β5 (4708S) (Cell Signalling Technology); anti-integrin αv (LM142) (Merck Millipore); anti-pan-talin (1 and 2) (53.8) (BioRad); anti-talin 1 (TA205) (Santa Cruz); anti-talin 2 (68E7) (Abcam); anti-integrin αVβ3 (LM609) (Abcam); anti-integrin β3 (AP3) (Abcam); anti-integrin β1 (LM534) (Millipore); anti-vinculin (hVIN-1) (Sigma Aldrich); anti-vinculin (V9131) (Sigma-Aldrich); anti-intersectin 1 (HPA018007) (Atlas Antibodies, Sigma-Aldrich); anti-NUMB (2733) (Cell Signaling Technologies); anti-EPS15L1 (HPA055309) (Atlas Antibodies, Sigma-Aldrich); anti-HIP1 (HPA013606) (Atlas Antibodies, Sigma-Aldrich); anti-WASL (HPA005750) (Atlas Antibodies, Sigma-Aldrich); anti-DAB2 (12906) (Cell Signaling Technologies); anti-paxillin (5H11) (Sigma Aldrich); anti-FAK (BD Biosciences); anti-zyxin (H-200) (Santa Cruz); anti-kindlin 2 (ab74030) (Abcam); anti-ICAP1 (115228) (Abcam); anti-DOK1 (HPA048561) (Atlas Antibodies, Sigma-Aldrich); polyclonal (rabbit) anti-phosphotyrosine (1000) (Cell Signaling); anti-cytokeratin (27988) (Abcam); anti-beta tubulin (DM1A) (Thermo Fisher Scientific); anti-vimentin (8978) (Abcam).

Techniques: Labeling, Binding Assay, Expressing

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Anti-inflammation conferred by stimulation of CD200R1 via Dok1 pathway in rat microglia after germinal matrix hemorrhage

doi: 10.1177/0271678X17725211

Figure Lengend Snippet: The adverse effects of silencing endogenous CD200R1 or Dok1 by CD200R1 siRNA or Dok1 siRNA at 24 h after GMH. (a) Representative western blot bands. (b) Quantitative analysis of CD200 showed that protein level of CD200 increased following the treatment with CD200Fc. (c) Quantitative analysis of CD200R1 showed CD200R1 increased in the treatment group and decreased in GMH group. CD200R1 siRNA intervention significantly decreased CD200R1 level. However, CD200R1 level does not change with Dok1 siRNA intervention. (d) Quantitative analysis of Dok1 showed that Dok1 expression significantly decreased by both Dok1 siRNA and CD200R1 siRNA intervention. Data are expressed as mean ± SD, *P < 0.05 vs Sham, #P < 0.05 vs vehicle, &P < 0.05 vs Treatment, n = 6/group, one-way ANOVA followed by the Tukey test.

Article Snippet: CD200R1 small interfering RNA (siRNA, 500 pmol, MyBioSource), scrambled siRNA (500 pmol, MyBioSource), and Dok1 siRNA (500 pmol, MyBioSource) in 0.5 μl PBS was infused at 1.0 μl/min through the Hamilton syringe at 24 h prior to GMH induction.

Techniques: Western Blot, Expressing

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Anti-inflammation conferred by stimulation of CD200R1 via Dok1 pathway in rat microglia after germinal matrix hemorrhage

doi: 10.1177/0271678X17725211

Figure Lengend Snippet: Effects of CD200R1 siRNA and Dok1 siRNA on IL-1beta and TNF-alpha expression in the presence of CD200Fc (1.5 mg/kg) at 24 h after GMH. (a) Representative western blots bands. (b) Quantitative analysis of IL-1beta. GMH enhanced while CD200Fc reduced the protein level of IL-1beta. (c) Quantitative analysis of TNF-alpha. GMH enhanced protein level of TNF-alpha while CD200Fc attenuated this induction. Data are expressed as mean ± SD, *P < 0.05 vs Sham, #P < 0.05 vs GMH + Vehicle, &P < 0.05 vs Treatment, n = 6/group, one-way ANOVA followed by the Tukey test.

Article Snippet: CD200R1 small interfering RNA (siRNA, 500 pmol, MyBioSource), scrambled siRNA (500 pmol, MyBioSource), and Dok1 siRNA (500 pmol, MyBioSource) in 0.5 μl PBS was infused at 1.0 μl/min through the Hamilton syringe at 24 h prior to GMH induction.

Techniques: Expressing, Western Blot

Journal: Oncoimmunology

Article Title: Docking protein-1 promotes inflammatory macrophage signaling in gastric cancer

doi: 10.1080/2162402X.2019.1649961

Figure Lengend Snippet:

Article Snippet: Full-length (FL) DOK1 cDNA (aa 1–481; 62 kDa) was inserted into pTarget (Promega, Madison, WI).

Techniques: Virus, Suspension