phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416
    Phospho Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho src family tyr416
    (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at <t>Tyr416</t> were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.
    Rabbit Anti Phospho Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Pip5k1γ regulates axon formation by limiting Rap1 activity"

    Article Title: Pip5k1γ regulates axon formation by limiting Rap1 activity

    Journal: Life Science Alliance

    doi: 10.26508/lsa.202302383

    (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at Tyr416 were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.
    Figure Legend Snippet: (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at Tyr416 were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.

    Techniques Used: Immunoprecipitation, Western Blot, Pull Down Assay, Binding Assay, Cell Culture, Molecular Weight, Staining, Fluorescence

    Reagents and Tools table.
    Figure Legend Snippet: Reagents and Tools table.

    Techniques Used: Recombinant

    phospho src family tyr416 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416 antibody
    NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src <t>(Tyr416)</t> and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).
    Phospho Src Family Tyr416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Newcastle disease virus activates diverse signaling pathways via Src to facilitate virus entry into host macrophages"

    Article Title: Newcastle disease virus activates diverse signaling pathways via Src to facilitate virus entry into host macrophages

    Journal: Journal of Virology

    doi: 10.1128/jvi.01915-23

    NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src (Tyr416) and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).
    Figure Legend Snippet: NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src (Tyr416) and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).

    Techniques Used: Adsorption, Flow Cytometry, Western Blot

    Antibodies used in this study
    Figure Legend Snippet: Antibodies used in this study

    Techniques Used:

    phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416
    Phospho Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416
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    phospho src family tyr416  (Cell Signaling Technology Inc)


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    rabbit anti phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho src family tyr416

    Rabbit Anti Phospho Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Protocol to characterize extracellular c-Src tyrosine kinase function through substrate interaction and phosphorylation"

    Article Title: Protocol to characterize extracellular c-Src tyrosine kinase function through substrate interaction and phosphorylation

    Journal: STAR Protocols

    doi: 10.1016/j.xpro.2023.102755


    Figure Legend Snippet:

    Techniques Used: Recombinant, Concentration Assay, Western Blot, Modification, Protease Inhibitor, Staining, Saline, Bradford Assay, Software, Electrophoresis

    phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416
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    phospho src family tyr416  (Cell Signaling Technology Inc)


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    phospho src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src family tyr416
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    Cell Signaling Technology Inc phospho src family tyr416
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    Cell Signaling Technology Inc rabbit anti phospho src family tyr416
    (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at <t>Tyr416</t> were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.
    Rabbit Anti Phospho Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho src family tyr416 antibody
    NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src <t>(Tyr416)</t> and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).
    Phospho Src Family Tyr416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at Tyr416 were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.

    Journal: Life Science Alliance

    Article Title: Pip5k1γ regulates axon formation by limiting Rap1 activity

    doi: 10.26508/lsa.202302383

    Figure Lengend Snippet: (A) Cortical neurons were treated with vehicle (Veh, DMSO) or UNC3230 (UNC) as indicated. Src family kinases phosphorylated at Tyr416 were immunoprecipitated and analyzed by Western blot with an anti-Fyn antibody as indicated. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (A, B) Normalized ratio of phospho-Tyr416-Fyn to total Fyn is shown (n = 3 independent experiments that are biological replicates). (C) Cortical neurons were treated as in (A), and the amount of active GTP-bound Rap1 was quantified by a pull-down assay using bacterially expressed GST-RBD (Ras/Rap-binding domain) and analyzed by Western blot with the indicated antibodies. The levels of phospho-Tyr515-C3G and total C3G were analyzed from lysates of cultured neurons. Detection of Tuj1 confirmed the analysis of comparable amounts of lysate. (C, D, E) Normalized ratio of active to total Rap1 (D) and phospho-Tyr515-C3G to total C3G (E) is shown (n = 3 independent experiments that are biological replicates). The molecular weight is indicated in kD. (F, H) Hippocampal neurons were treated with vehicle (DMSO) or UNC3230 for 5 h at 2 d.i.v. and stained with anti-phospho-Tyr515-C3G (green) and anti-C3G (magenta) (F) or with anti-GTP-Rap1 (red) and anti-Rap1 (cyan, total Rap1) (G) antibodies. Red arrows mark the neurite with the strongest signal, whereas yellow arrows mark the other neurites. (F, G, H, I) Polarization index was calculated for (F, H) as the fluorescence intensity (a.u.) of the brightest neurite divided by the average fluorescence intensity (a.u.) of the other minor neurites of the same cell. Scale bar: 20 μm. Values are means ± SEM. (B, D, E, G, I) : unpaired t test. Source data are available for this figure.

    Article Snippet: Rabbit anti-phospho-Src family (Tyr416) , Cell Signaling Technology , Cat# 6943; RRID: AB_10013641.

    Techniques: Immunoprecipitation, Western Blot, Pull Down Assay, Binding Assay, Cell Culture, Molecular Weight, Staining, Fluorescence

    Reagents and Tools table.

    Journal: Life Science Alliance

    Article Title: Pip5k1γ regulates axon formation by limiting Rap1 activity

    doi: 10.26508/lsa.202302383

    Figure Lengend Snippet: Reagents and Tools table.

    Article Snippet: Rabbit anti-phospho-Src family (Tyr416) , Cell Signaling Technology , Cat# 6943; RRID: AB_10013641.

    Techniques: Recombinant

    NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src (Tyr416) and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).

    Journal: Journal of Virology

    Article Title: Newcastle disease virus activates diverse signaling pathways via Src to facilitate virus entry into host macrophages

    doi: 10.1128/jvi.01915-23

    Figure Lengend Snippet: NDV induces tyrosine phosphorylation of Src for entry into HD11 cells. (A and B) Treatment with Genistein inhibited internalization of NDV. HD11 cells were treated with Genistein or DMSO and then subjected to adsorption (A) and internalization (B) assays. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. (C) Y416 phosphorylation of Src was induced by NDV during the entry process. HD11 cells were either inoculated with NDV at an MOI of 10 or mock inoculated. At each of the indicated time point, cell lysates were analyzed by Western blotting using antibodies against p-Src (Tyr416) and Src, respectively. GAPDH was used as a control. The relative intensity of p-Src (Tyr416) and Src were normalized to GAPDH. (D) Y416 phosphorylation of Src was induced by NDV in HD11 cells and primary chicken macrophage, rather than DF-1 cells. HD11 cells, primary chicken macrophage and DF-1 cells were either inoculated with NDV at an MOI of 5 and 10 or mock inoculated. At 30 mpi with NDV, the levels of p-Src (Tyr416) and Src by Western blotting as described in Fig. 3C. (E–H) Treatment with Src inhibitors inhibited the internalization of NDV. HD11 cells were pretreated with Saracatinib (E and F) and Dasatinib (G and H), or mock-treated with DMSO, after which adsorption and internalization assays were performed. (I) Treatment with Saracatinib inhibited NDV-induced phosphorylation of Src. HD11 cells were pretreated with either DMSO or Saracatinib. The cells were inoculated or mock inoculated with NDV. At 30 mpi, the levels of p-Src and Src were determined by Western blotting. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).

    Article Snippet: Phospho-Src , Phospho-Src Family (Tyr416) antibody , CST , 2101.

    Techniques: Adsorption, Flow Cytometry, Western Blot

    Antibodies used in this study

    Journal: Journal of Virology

    Article Title: Newcastle disease virus activates diverse signaling pathways via Src to facilitate virus entry into host macrophages

    doi: 10.1128/jvi.01915-23

    Figure Lengend Snippet: Antibodies used in this study

    Article Snippet: Phospho-Src , Phospho-Src Family (Tyr416) antibody , CST , 2101.

    Techniques: