Journal: The Journal of biological chemistry

Article Title: Activation of p38 mitogen-activated protein kinase by c-Abl-dependent and -independent mechanisms.

doi: 10.1074/jbc.271.39.23775

Figure Lengend Snippet: FIG. 1. Activation of p38 MAPK by CDDP. NIH3T3 cells were treated with 50 mM CDDP for the indicated times (A) or with the indicated concentrations of CDDP for 3 h (B). Total cell lysates were immunoprecipitated with anti-p38 MAPK antibody, and in vitro immune complex kinase reactions were performed using GST-ATF2 fusion protein as substrate. The proteins were separated by 10% SDS-PAGE, stained with Coomassie Blue (second panels, A and B) and analyzed by autoradiography (top panels, A and B). Anti-p38 MAPK immunoprecipitates were also analyzed by immunoblotting with anti-p38 (third panels, A and B) or anti-Tyr(P) (fourth panels, A and B) antibodies. The fold increase in p38 MAPK activity is shown (bottom panels, A and B) as the mean (bars, S.E.) for three independent experiments.

Article Snippet: Lysates were incubated with anti-SAP kinase (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-p38 MAPK (16) antibodies for 1 h at 4 °C and then for 45 min after the addition of protein A-Sepharose.

Techniques: Activation Assay, Immunoprecipitation, In Vitro, Immune Complex Kinase Assay, SDS Page, Staining, Autoradiography, Western Blot, Activity Assay

Journal: The Journal of biological chemistry

Article Title: Activation of p38 mitogen-activated protein kinase by c-Abl-dependent and -independent mechanisms.

doi: 10.1074/jbc.271.39.23775

Figure Lengend Snippet: FIG. 2. Activation of p38 MAPK by different DNA-damaging agents in diverse cell types. A, U-937 cells were treated with 50 mM CDDP for 3 h. B, U-937 cells were treated with 50 mM ara-C for the indicated times. Total cell lysates were immunoprecipitated with anti- p38 MAP kinase antibody, and in vitro immune complex kinase reac- tions containing GST-ATF2 fusion protein were analyzed by 10% SDS-PAGE and autoradiography (upper panel). Anti-p38 MAPK immunoprecipitates from control and ara-C-treated cell lysates were also analyzed by immunoblotting with anti-Tyr(P) antibodies (lower panel). C, NIH3T3 cells were treated with 50 mM ara-C for the indicated times. The p38 MAPK activity was measured as described above. D, U-937 cells were treated with 20 gray IR and harvested at the indicated times. Total cell lysates were immunoprecipitated with either anti-p38 MAPK or anti-SAPK antibodies. In vitro immune complex kinase reac- tions containing GST-ATF2 (upper panel) or GST-Jun (bottom panel) fusion proteins were analyzed by 10% SDS-PAGE and autoradiography. E, NIH3T3 cells were treated with 50 mM CDDP for 3 h, 1 mM MMS for 3 h, or 80 J/m2 UV (harvested at 30 min). The p38 MAPK activity was measured as described above. The fold activation in kinase activities is shown at the bottom.

Article Snippet: Lysates were incubated with anti-SAP kinase (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-p38 MAPK (16) antibodies for 1 h at 4 °C and then for 45 min after the addition of protein A-Sepharose.

Techniques: Activation Assay, Immunoprecipitation, In Vitro, Immune Complex Kinase Assay, SDS Page, Autoradiography, Control, Western Blot, Activity Assay

Journal: The Journal of biological chemistry

Article Title: Activation of p38 mitogen-activated protein kinase by c-Abl-dependent and -independent mechanisms.

doi: 10.1074/jbc.271.39.23775

Figure Lengend Snippet: FIG. 3. c-Abl-dependent activation of p38 MAPK in response to CDDP. A, NIH3T3, Abl2/2 and Abl1 cells were treated with 100 mM CDDP for 3 h. Total cell lysates were immunoprecipitated with anti-p38 MAPK antibody, and in vitro immune complex kinase reactions con- taining GST-ATF2 fusion protein were analyzed by 10% SDS-PAGE and autoradiography (left panel). The fold increase in activity is shown at the bottom. Total cell lysates from NIH3T3, Abl2/2, and Abl1 cells were also immunoprecipitated with anti-Abl antibody. Protein precipi- tates were analyzed by immunoblotting with anti-Abl (right panel). B, Abl2/2 cells were treated with 100 mM CDDP for the indicated times. NIH3T3 cells were treated with 100 mM CDDP for 3 h as a positive control. p38 MAPK activity (upper panel) was assayed as described above. The fold increase in p38 MAPK activity is shown at the bottom panel (one of the three representative experiments is shown). C, NIH3T3 and Abl2/2 cells were treated with 50 mM ara-C for 3 h. Abl2/2

Article Snippet: Lysates were incubated with anti-SAP kinase (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-p38 MAPK (16) antibodies for 1 h at 4 °C and then for 45 min after the addition of protein A-Sepharose.

Techniques: Activation Assay, Immunoprecipitation, In Vitro, Immune Complex Kinase Assay, SDS Page, Autoradiography, Activity Assay, Western Blot, Positive Control

Journal: The Journal of biological chemistry

Article Title: Activation of p38 mitogen-activated protein kinase by c-Abl-dependent and -independent mechanisms.

doi: 10.1074/jbc.271.39.23775

Figure Lengend Snippet: FIG. 4. Abl-dependent and -independent activation of p38 MAPK and SAP/JNK kinases by CDDP, MMS, or UV. NIH3T3 and Abl2/2 cells were treated with 1 mM MMS for 3 h (left panels), 100 mM CDDP for 3 h (left panels), or 80 J/m2 UV (harvested at 15 min) (right panels). Total cell lysates were immunoprecipitated with anti-p38 MAPK (A) or anti-SAPK (B) antibodies, and in vitro immune complex kinase reactions containing GST-ATF2 (A) or GST-Jun (B) fusion pro- teins were analyzed by 10% SDS-PAGE and autoradiography.

Article Snippet: Lysates were incubated with anti-SAP kinase (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-p38 MAPK (16) antibodies for 1 h at 4 °C and then for 45 min after the addition of protein A-Sepharose.

Techniques: Activation Assay, Immunoprecipitation, In Vitro, Immune Complex Kinase Assay, SDS Page, Autoradiography