Journal:

Article Title: New functional roles for non-collagenous domains of basement membrane collagens

doi:

Figure Lengend Snippet: Functions of NC1 and NC1 fragments of collagen IV, XV and XVIII

Article Snippet: , FAK and PI3K phosphorylation (α3) ( Pasco et al., 2000a ) , .

Techniques: Inhibition, Chemotaxis Assay, Phospho-proteomics, Activation Assay, Migration, In Vitro, Membrane, Angiogenesis Assay

Journal:

Article Title: New functional roles for non-collagenous domains of basement membrane collagens

doi:

Figure Lengend Snippet: Biological activities of immobilized versus soluble NC1 and endostatin fragments. Immobilized NC1 domains from collagen IV, XV and XVIII induce proliferation, survival and migration of different cell types. These effects correlate with an increase in PI3K, FAK and paxillin phosphorylation. By contrast, soluble NC1 or endostatin fragments bind to various receptors on the surface of the cells and decrease phosphorylation of PI3K, FAK and paxillin. Endostatin as well as NC1(IV) induces a decrease in the transcription of different genes. Endostatin interacts with VEGF-R2 and thus decreases the binding of VEGF to its receptor; endostatin also interacts directly with MMP-2, inhibiting the activation of the enzyme. Taken together, these soluble fragments act in the opposite way to their original molecules, negatively regulating the proliferation and the migration of different cell types and inducing apoptosis and extracellular matrix disorganization.

Article Snippet: , FAK and PI3K phosphorylation (α3) ( Pasco et al., 2000a ) , .

Techniques: Migration, Phospho-proteomics, Binding Assay, Activation Assay

Journal: Oncology Letters

Article Title: Diallyl disulfide effect on the invasion and migration ability of HL-60 cells with a high expression of DJ-1 in the nucleus through the suppression of the Src signaling pathway

doi: 10.3892/ol.2018.8139

Figure Lengend Snippet: The expression of p-Src and p-Fak in the HL-60 cells with a high expression of DJ-1 in the nucleus induced by diallyl disulfide for 0, 5, 15, 30 or 60 min. (A) A representative western blot analysis presenting the expression of p-Src and p-Fak. (B) Comparisons between the expression levels of p-Src and p-Fak. *P<0.05 vs. the control. p-Src, phosphorylated Src protein; p-Fak, phosphorylated Fak protein; DJ-1, parkinsonism associated deglycase.

Article Snippet: Phosphorylated Src (p-Src) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA; cat. no. 2101; 1:1,000 dilution) and phosphorylated Fak (p-Fak) was purchased from Abgent, Inc. (cat no. AJ1285e; 1:1,000 dilution). β-actin monoclonal antibodies were purchased from Abgent, Inc. (cat. no. AM1021B-ev; 1:2,000 dilution).

Techniques: Expressing, Western Blot

Journal: Oncology Letters

Article Title: Diallyl disulfide effect on the invasion and migration ability of HL-60 cells with a high expression of DJ-1 in the nucleus through the suppression of the Src signaling pathway

doi: 10.3892/ol.2018.8139

Figure Lengend Snippet: DADS affects the Src signaling pathway in a control group, empty vector group and high expression group. (A) Protein expression of p-Src, Src, P-Fak, Fak and integrin was examined using western blot analysis. (B) Quantified protein expression of p-Src and p-Fac across the three groups. *P<0.05 vs. the control. DADS, diallyl disulfide; DJ-1, parkinsonism associated deglycase; p-Src, phosphorylated Src protein; p-Fak, phosphorylated Fak protein.

Article Snippet: Phosphorylated Src (p-Src) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA; cat. no. 2101; 1:1,000 dilution) and phosphorylated Fak (p-Fak) was purchased from Abgent, Inc. (cat no. AJ1285e; 1:1,000 dilution). β-actin monoclonal antibodies were purchased from Abgent, Inc. (cat. no. AM1021B-ev; 1:2,000 dilution).

Techniques: Plasmid Preparation, Expressing, Western Blot

Journal: Oncology Letters

Article Title: Diallyl disulfide effect on the invasion and migration ability of HL-60 cells with a high expression of DJ-1 in the nucleus through the suppression of the Src signaling pathway

doi: 10.3892/ol.2018.8139

Figure Lengend Snippet: DADS and Src inhibitor inhibit the Src signaling pathway in the control group, the empty vector group and the high expression group. Protein expression of p-Src, Src, P-Fak, Fak and integrin was examined by (A) western blot analysis and (B) quantitative analysis. *P<0.05 vs. the control. DADS, diallyl disulfide; p-Src, phosphorylated Src protein; p-Fak, phosphorylated Fak protein; DJ-1, parkinsonism associated deglycase.

Article Snippet: Phosphorylated Src (p-Src) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA; cat. no. 2101; 1:1,000 dilution) and phosphorylated Fak (p-Fak) was purchased from Abgent, Inc. (cat no. AJ1285e; 1:1,000 dilution). β-actin monoclonal antibodies were purchased from Abgent, Inc. (cat. no. AM1021B-ev; 1:2,000 dilution).

Techniques: Plasmid Preparation, Expressing, Western Blot

Journal: Nature cell biology

Article Title: Integrin endosomal signalling suppresses anoikis

doi: 10.1038/ncb3250

Figure Lengend Snippet: a, Representative immunoblot of subcellular fractionation of FAK+/+ and FAK-/- MEFs (five independent experiments). b, Representative immunoblot of recombinant FAK (phosphorylated/activated or non-activated: n.a) recruitment to insoluble endosomal pellet (P) and soluble supernatant (S) fractions isolated from FAK -/- MEFs (five independent experiments). c, Representative immunoblot analysing the recruitment of recombinant FAK to purified endosomes derived from either control- or β1-integrin-silenced FAK -/- MEFs (five independent experiments). d, Representative immunoblot analysing the activation of recombinant FAK in purified endosome fractions derived from FAK -/- MEFs in the presence or absence of 10 µM ATP (five independent experiments). e, Subcellular fractionation of FAK+/+ MEFs transfected with GFP-FAK FAT or GFP-FAK FERM. Shown are representative immunoblots of GFP-FAK fragments and endogenous FAK (total-FAK) from two independent experiments. f, MDA-MB-231 cells plated on 20 μm round fibronectin-coated micropatterns (45 min) ± dynasore. Representative maximum projections and quantification of pFAK (mean fluorescence ± SEM, n=3 independent experiments, 10 cells analysed / experiment). Lys: cell lysate; PM: plasma membrane; Cyto: cytosol; Endo: Endosomal fraction. Student's two-tailed unpaired t-test P =0.01. Uncropped images of blots are shown in supplementary figure 9.

Article Snippet: Therefore, the endosomal fraction derived from FAK -/- MEFs was resuspended in buffer containing 20 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM MgCl 2 , 1 mM DTT and protease & phosphatase inhibitors and subsequently incubated with 0.5 μg of either phosphorylated or non-phosphorylated FAK (ProQinase) or GST for 2 h at room temperature.

Techniques: Western Blot, Fractionation, Recombinant, Isolation, Purification, Derivative Assay, Control, Activation Assay, Transfection, Fluorescence, Clinical Proteomics, Membrane, Two Tailed Test

Journal: PLoS ONE

Article Title: Neuregulin Facilitates Nerve Regeneration by Speeding Schwann Cell Migration via ErbB2/3-Dependent FAK Pathway

doi: 10.1371/journal.pone.0053444

Figure Lengend Snippet: The physical association of erbB-FAK signaling was analyzed by the ratio expressed as phosphorylated FAK (p-FAK) over total FAK. Note that NRG β1 extensively increases erbB-FAK activation in both PLL and laminin-coated plates with the most significant effect observed in the later group. * P <0.05 as compared to that of control value.

Article Snippet: For detection of erbB2, erbB3 and its downstream signaling molecules, anti-phospho-tyrosine antibody 4G10 (Upstate Biotechnology, Lake Placid, NY, USA), anti-erbB2, anti-erbB3 (Santa Cruz, CA, USA), anti-FAK (Biosources, Nivelles, Belgium), anti-phosphorylated FAK (p-FAK, Biosources, Nivelles, Belgium), and anti-β-actin (BD Pharmingen, San Jose, CA, USA) primary antibodies were applied to the nitrocellulose membranes.

Techniques: Activation Assay

Journal: PLoS ONE

Article Title: Neuregulin Facilitates Nerve Regeneration by Speeding Schwann Cell Migration via ErbB2/3-Dependent FAK Pathway

doi: 10.1371/journal.pone.0053444

Figure Lengend Snippet: Following PNI [as created by end-to-side neurorraphy (ESN)], NRG would activate the erbB2-FAK pathway that promotes the Schwann cells to migrate to the distal stump for successful axonal guidance and nerve regeneration. Un: ulnar nerve; Mc: musculocutaneous nerve; ECM: extracellular matrix.

Article Snippet: For detection of erbB2, erbB3 and its downstream signaling molecules, anti-phospho-tyrosine antibody 4G10 (Upstate Biotechnology, Lake Placid, NY, USA), anti-erbB2, anti-erbB3 (Santa Cruz, CA, USA), anti-FAK (Biosources, Nivelles, Belgium), anti-phosphorylated FAK (p-FAK, Biosources, Nivelles, Belgium), and anti-β-actin (BD Pharmingen, San Jose, CA, USA) primary antibodies were applied to the nitrocellulose membranes.

Techniques: