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: Neuroscience

Article Title: Linkage of non-receptor tyrosine kinase Fyn to mGlu5 receptors in striatal neurons in a depression model

doi: 10.1016/j.neuroscience.2020.02.048

Figure Lengend Snippet: (A) Effects of social isolation on pY416, Fyn, and Src levels in the CPu. (B) Effects of social isolation on pY416, Fyn, and Src levels in the NAc. Note that social isolation (SI) had no significant effect on phosphorylation and expression of Fyn and Src in the CPu (A) and NAc (B) as compared to control (Con) rats. Representative immunoblots are shown left to the quantified data. Data were statistically analyzed using Student’s t-test (n = 6 per group) with ‘n’ equal to the number of animals.

Article Snippet: Mouse antibodies include those against Fyn (RRID:AB_627642; Santa Cruz), Src (RRID:AB_2106058; Cell Signaling), mGlu5 (RRID:AB_1523943; Abcam, Cambridge, MA), and transferrin receptors (TfR, RRID:AB_2533029; ThermoFisher).

Techniques: Isolation, Expressing, Western Blot

Journal: Neuroscience

Article Title: Linkage of non-receptor tyrosine kinase Fyn to mGlu5 receptors in striatal neurons in a depression model

doi: 10.1016/j.neuroscience.2020.02.048

Figure Lengend Snippet: (A) Effects of social isolation on Y416 phosphorylation of immunoprecipitated Fyn proteins. (B) Effects of social isolation on Y416 phosphorylation of immunoprecipitated Src proteins. Fyn and Src were precipitated from the striatum of socially isolated (SI) rats and control rats by immunoprecipitation (IP). Immunoprecipitated proteins were visualized by immunoblots (IB) with indicated antibodies. Representative immunoblots are shown left to the quantified data. Data were statistically analyzed using Student’s t-test (n = 6 per group) with ‘n’ equal to the number of animals.

Article Snippet: Mouse antibodies include those against Fyn (RRID:AB_627642; Santa Cruz), Src (RRID:AB_2106058; Cell Signaling), mGlu5 (RRID:AB_1523943; Abcam, Cambridge, MA), and transferrin receptors (TfR, RRID:AB_2533029; ThermoFisher).

Techniques: Isolation, Immunoprecipitation, Western Blot

Journal: Neuroscience

Article Title: Linkage of non-receptor tyrosine kinase Fyn to mGlu5 receptors in striatal neurons in a depression model

doi: 10.1016/j.neuroscience.2020.02.048

Figure Lengend Snippet: (A) Effects of social isolation on Fyn kinase activity. (B) Effects of social isolation on Src kinase activity. Fyn and Src were immunoprecipitated from the striatum of socially isolated (SI) rats and control rats. Data were statistically analyzed using Student’s t-test (n = 6 per group) with ‘n’ equal to the number of animals.

Article Snippet: Mouse antibodies include those against Fyn (RRID:AB_627642; Santa Cruz), Src (RRID:AB_2106058; Cell Signaling), mGlu5 (RRID:AB_1523943; Abcam, Cambridge, MA), and transferrin receptors (TfR, RRID:AB_2533029; ThermoFisher).

Techniques: Isolation, Activity Assay, Immunoprecipitation

Journal: Neuroscience

Article Title: Linkage of non-receptor tyrosine kinase Fyn to mGlu5 receptors in striatal neurons in a depression model

doi: 10.1016/j.neuroscience.2020.02.048

Figure Lengend Snippet: (A) Coimmunoprecipitation (IP) of Fyn/Src and mGlu5 receptors in striatal neurons as detected with an anti-mGlu5 antibody (Ab). (B) Reverse coimmunoprecipitation of Fyn and mGlu5 receptors in striatal neurons as detected with an anti-Fyn antibody. Note that Fyn, Src, and mGlu5 receptors were seen in mGlu5 precipitates in lane 5 (L5) (A). No specific bands were shown in lanes 3 and 4 due to the absence of a precipitating antibody (L3) and the presence of an irrelevant IgG (L4). Solubilized rat striatal lysates were used in IP assays. Immunoprecipitated proteins were visualized by immunoblots (IB) with indicated antibodies.

Article Snippet: Mouse antibodies include those against Fyn (RRID:AB_627642; Santa Cruz), Src (RRID:AB_2106058; Cell Signaling), mGlu5 (RRID:AB_1523943; Abcam, Cambridge, MA), and transferrin receptors (TfR, RRID:AB_2533029; ThermoFisher).

Techniques: Immunoprecipitation, Western Blot

Journal: Neuroscience

Article Title: Linkage of non-receptor tyrosine kinase Fyn to mGlu5 receptors in striatal neurons in a depression model

doi: 10.1016/j.neuroscience.2020.02.048

Figure Lengend Snippet: (A) Effects of social isolation (SI) on the Fyn-mGlu5 interaction. (B) Effects of social isolation on SFK Y416 phosphorylation in mGlu5 precipitates. (C) Effects of social isolation on the Src-mGlu5 interaction. Note that social isolation elevated the Fyn-mGlu5 interaction (A), although the Src-mGlu5 interaction was not significantly altered (C). Representative immunoblots are shown left to the quantified data. Solubilized rat striatal lysates were used in immunoprecipitation (IP). Immunoprecipitated proteins were visualized by immunoblots (IB) with indicated antibodies. Data were statistically analyzed using Student’s t-test (n = 6 per group). *P < 0.05 versus double-housed control animals. P values = 0.009 (A), 0.008 (B), and 0.406 (C).

Article Snippet: Mouse antibodies include those against Fyn (RRID:AB_627642; Santa Cruz), Src (RRID:AB_2106058; Cell Signaling), mGlu5 (RRID:AB_1523943; Abcam, Cambridge, MA), and transferrin receptors (TfR, RRID:AB_2533029; ThermoFisher).

Techniques: Isolation, Western Blot, Immunoprecipitation