Journal: International journal of oncology

Article Title: Synergistic cell growth inhibition by the combination of amrubicin and Akt-suppressing tyrosine kinase inhibitors in small cell lung cancer cells: implication of c-Src and its inhibitor.

doi: 10.3892/ijo_00000195

Figure Lengend Snippet: Figure 5. Effect of PP2 and dasatinib on Akt activity and synergistic interaction with amrubicin (AMR) in N417 cells. (A and C) N417 cells were treated with the indicated concentrations of PP2 or dasatinib for 5 h. c-Src was immunoprecipitated (IP) from 1 mg of total cellular protein by anti-c-Src and subjected to Western blot (WB) analysis with anti-phospho-Src (Tyr416) or c-Src. Total cellular protein (50 μg) from the same cell lysate was subjected to Western blot analysis with anti-phospho-Akt (p-Akt), anti-Akt or anti-ß-actin antibody. (B and D) N417 cells were treated by a combination of PP2 or dasatinib with AMR for 72 h. Dose-response curves were plotted on the basis of the data derived from MTT assay. The survival cell fraction was expressed as the relative optical density (OD) in reference to that of the untreated cells. The combination effects were evaluated with isobologram analysis. The envelopes of additivity are defined by three isoeffect lines constructed from the dose-response curves of the single agents. The concentration producing 50% cell growth inhibition (IC50) of PP2, dasatinib or AMR alone is expressed as 1 on the ordinate or the abscissa. The plotted data points show the relative values of the concentrations producing IC50.

Article Snippet: Antibody against Akt, phospho-Akt (Ser473), c-Kit, insulin-like growth factor I receptor (IGF-IR) and phospho-Src-family (Tyr416) were purchased from Cell Signaling Technology.

Techniques: Activity Assay, Immunoprecipitation, Western Blot, Derivative Assay, MTT Assay, Construct, Concentration Assay, Inhibition

Journal: Cancers

Article Title: Rac1 Signaling: From Intestinal Homeostasis to Colorectal Cancer Metastasis

doi: 10.3390/cancers12030665

Figure Lengend Snippet: Upstream effectors; RhoGAsP, RhoGDIs, GEFs.

Article Snippet: PLCG1, PLC-gamma-1 , Phospholipase C Gamma 1 , Hydrolysis of phosphatidylinositol 4,5-bisphosphate to 1,4,5-trisphosphate (IP3) and diacylglycerol , Critical for EGF-induced Rac1 activation in vivo. PLC-g SH3 domain acts as a Rac1 guanine nucleotide exchange in vivo. Cytoskeleton remodeling and cell migration [ ]. , .

Techniques: Activity Assay, Migration, Membrane, Binding Assay, Over Expression, Microarray, Activation Assay, Expressing, Adhesive, Mutagenesis, Genome Wide, Variant Assay, Blocking Assay, In Vivo

Journal: Cancers

Article Title: Rac1 Signaling: From Intestinal Homeostasis to Colorectal Cancer Metastasis

doi: 10.3390/cancers12030665

Figure Lengend Snippet: Interacting partners and functional consequences.

Article Snippet: PLCG1, PLC-gamma-1 , Phospholipase C Gamma 1 , Hydrolysis of phosphatidylinositol 4,5-bisphosphate to 1,4,5-trisphosphate (IP3) and diacylglycerol , Critical for EGF-induced Rac1 activation in vivo. PLC-g SH3 domain acts as a Rac1 guanine nucleotide exchange in vivo. Cytoskeleton remodeling and cell migration [ ]. , .

Techniques: Functional Assay, Membrane, Activity Assay, Activation Assay, Mutagenesis, Over Expression, Binding Assay, Migration, Translocation Assay, Expressing, Transduction, Scaffolding, Control, Sequencing, Extraction, Inhibition, Knockdown

Journal: Frontiers in Immunology

Article Title: Tumor Cell-Secreted ISG15 Promotes Tumor Cell Migration and Immune Suppression by Inducing the Macrophage M2-Like Phenotype

doi: 10.3389/fimmu.2020.594775

Figure Lengend Snippet: Interferon-stimulated gene 15 (ISG15)-induced CCL18 secretion by macrophages was dependent on the ISG15 receptor, leukocyte function-associated antigen-1 (LFA-1), and SRC family kinase (SFK) signaling. (A) Immunofluorescence demonstrated that the ISG15 receptor CD11a/CD18, also called LFA-1, was expressed on the membranes of human macrophages. Macrophages were treated with 295nM rISG15 for 24 h. Scale bars, 10 μm. (B) ELISA results showed small molecular inhibitors of LFA-1, A286982, inhibited CCL18 secretion by macrophages. (C) Small molecular inhibitors of LFA-1 and SRC, A286982 and PP2, hindered the ISG15-induced activation of SFK signaling. (D) PP2 reduced the CCL18 secretion by ISG15-treated macrophages in a dose-dependent manner, as determined by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Protein extracts were resolved by 10% SDS–PAGE, transferred to PVDF membranes (Roche), and probed with antibodies directed against human ISG15 (1:1,000; Abnova, catalog no. A155801), SRC (1:1,000; CST, catalog no. 9935T), and β-actin (1:3,000; Abcam, catalog no. ab69512).

Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay, Activation Assay