Journal: Molecular cell

Article Title: MRI is a DNA Damage Response Adaptor during Classical Non-Homologous End Joining

doi: 10.1016/j.molcel.2018.06.018

Figure Lengend Snippet: (A) Western blot analyses of DDR proteins that co-immunoprecipitated with FLAG-HA-tagged MRI in WT and Ku70−/− abl pre-B cell nuclear extracts using anti-HA. (B) Schematic of sequential immunoprecipitations of MRI followed by Ku80 in cells expressing HA-MRI and FLAG-Ku80 (left). Western blot analyses of ATM, DNA-PKcs, Ku70, FLAG-Ku80, and HA-MRI from MRI−/− abl pre-B cell nuclear extract after first immunoprecipitation with anti-HA (Input HA IP) and following second immunoprecipitation with anti-FLAG or an IgG isotype control (right). (C) Western blot analyses of purified recombinant biotin-FLAG-ATM (bio-F-ATM) co-immunoprecipitated with 50 nM, 100 nM, or 200 nM of MBP-tagged human MRI, MRIΔN, and MRIΔC proteins. (D) Western blot analyses of DDR proteins that associated with FLAG-purified MRI in different-sized fractions separated by a sucrose gradient column.

Article Snippet: 40 nM recombinant biotin-FLAG-tagged human ATM was incubated with MBP-tagged human MRI, MRI ΔN , or MRI ΔC (50, 100, or 200 nM) and 100 ng bovine serum albumin (BSA, New England Biolabs) in A buffer (25 mM Tris-HCl pH 8.0, 100 mM NaCl, 10% v/v glycerol) at room temperature for 15 minutes in a final volume of 30 μL.

Techniques: Western Blot, Immunoprecipitation, Expressing, Purification, Recombinant

Journal: Molecular cell

Article Title: MRI is a DNA Damage Response Adaptor during Classical Non-Homologous End Joining

doi: 10.1016/j.molcel.2018.06.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 40 nM recombinant biotin-FLAG-tagged human ATM was incubated with MBP-tagged human MRI, MRI ΔN , or MRI ΔC (50, 100, or 200 nM) and 100 ng bovine serum albumin (BSA, New England Biolabs) in A buffer (25 mM Tris-HCl pH 8.0, 100 mM NaCl, 10% v/v glycerol) at room temperature for 15 minutes in a final volume of 30 μL.

Techniques: Blocking Assay, Recombinant, Cell Isolation, Magnetic Beads, Mass Spectrometry, Magnetic Resonance Imaging, Sequencing, Plasmid Preparation, Software, High Content Screening, Flow Cytometry, Inverted Microscopy, Laser-Scanning Microscopy, Spectrophotometry, Irradiation