Journal: bioRxiv

Article Title: BRD4 regulates Aurora B kinase activity

doi: 10.1101/2025.08.06.668960

Figure Lengend Snippet: A. Aurora B interaction with BRD4 is unaffected by its phosphorylation. Co-IP of Aurora B with Flag-BRD4 on Flag beads following a kinase assay with or without ATP. B. JNK activation and phosphorylation of BRD4 in vivo abrogates BRD4-Aurora B interaction. Immunoblots showing Aurora B co-immunoprecipitated by anti-BRD4 with total and T1212 phosphorylated BRD4 from HCT116 cells treated with or without JNK activator anisomycin. C. JNK activation increases MCAK Ser95 phosphorylation. Immunoblots showing MCAK pSer95 and pJNK levels in HCT 116 cells with or without JNK activation by anisomycin for increasing durations of time. Data is representative of two independent experiments. D. JNK inhibition decreases MCAK Ser95 phosphorylation. Immunoblots showing MCAK pSer95 levels in HCT116 cells with or without anisomycin and JNK inhibitor D-JNK1. E. JNK does not directly phosphorylate MCAK or affect Aurora B activity. Autoradiograph of kinase assays with equimolar Aurora B and active JNK with MCAK as substrate. F. MCAK phosphorylation is dependent on JNK phosphorylation of BRD4. Immunoblots showing MCAK pSer95 levels in DLD-BRD4-IAA7 cells transfected with or without WT or 3A-BRD4 and treated with or without auxin and anisomycin.

Article Snippet: Purified recombinant kinase active human GST-tagged Aurora B and JNK1 was purchased from SignalChem.

Techniques: Phospho-proteomics, Co-Immunoprecipitation Assay, Kinase Assay, Activation Assay, In Vivo, Western Blot, Immunoprecipitation, Inhibition, Activity Assay, Autoradiography, Transfection

Journal: Journal of Biological Chemistry

Article Title: Slit Protein-mediated Inhibition of CXCR4-induced Chemotactic and Chemoinvasive Signaling Pathways in Breast Cancer Cells

doi: 10.1074/jbc.m308083200

Figure Lengend Snippet: FIG. 8. Slit specifically inhibits CXCL12-induced p44/42 MAP kinase activity. Cell lysates were obtained from Slit (100 g/ml) or control (100 g/ml) pretreated DU4475 cells after stimulation with CXCL12 (10 nM). The lysates were immunoprecipitated with p44/42 (A), p38 MAP kinase (B), or JNK kinase (C) antibodies. The immune com- plex for p44/42 was run on SDS-PAGE and immunoblotted with phos- pho-p44/42 antibody (p-p44/42) (A, upper panel). The protein levels were monitored by stripping the blot and reprobing with anti-p44/42 antibodies (A, lower panel). The p38 MAP kinase and JNK immunopre- cipitates were subjected to in vitro kinase assay using myelin basic protein (MBP) and c-Jun, respectively, along with labeled ATP as substrates.

Article Snippet: Briefly, cell lysates were immunoprecipitated with JNK antibody (Santa Cruz Biotechnology).

Techniques: Activity Assay, Control, Immunoprecipitation, SDS Page, Stripping Membranes, In Vitro, Kinase Assay, Labeling