flag c myc ha  (MedChemExpress)

Journal: Molecular cell

Article Title: NBS1 phosphorylation status dictates repair choice of dysfunctional telomeres

doi: 10.1016/j.molcel.2017.01.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies phospho-CHK1 Cell Signaling Technology 2348 phospho-CHK2 BD Biosciences 611,570 γ-H2AX Millipore 05–636 phospho-NBS1 S432 Abcam ab12297 phospho-RPA32 (S4/S8) Bethyl A300-245A BARD1 (H-300) Santa Cruz sc-11,438 53BP1 Santa Cruz sc-22760 TRF2 Millipore 05–521 c-Myc (A14) Santa Cruz sc-789 GFP Santa Cruz Sc-9996 Flag Sigma-Aldrich F3165 HA Sigma-Aldrich H3663 PP1-α Santa Cruz sc-443 γ-tubulin (clone GTU-488) Sigma-Aldrich T6557 anti-Myc agarose beads Sigma-Aldrich A7470 anti-Flag agarose beads Sigma-Aldrich A2220 Biological Samples Chemicals, Peptides, and Recombinant Proteins Roscovitine Sigma-Aldrich R7772 Phosphatase inhibitor cocktail Sigma-Aldrich P 5726 1NM-PP1 Calbiochem 529581-1MG Thymidine Sigma-Aldrich T1895-5G Aphidicolin Sigma-Aldrich A4487-1ML FAM-labeled NBS1 peptides (hNBS1 423–438) This paper N/A FAM-labeled Apollo peptides (hAPOLLO 500–513) This paper Human TRF2 TRFH (residues 42-245) Chen et al., 2008b NBS1 WT and mutant peptides (423–438) Convenience Biology Critical Commercial Assays Site-directed mutagenesis Stratagene 200521 Deposited Data atomic coordinates of the TRF2-NBS1 complex This paper PDB: 5WQD Experimental Models: Cell Lines Human NBS-ILB1 cells Falck et al ., 2012 , EMBO Reports.

Techniques: Recombinant, Mutagenesis, Sequencing, Software

Journal: PLoS Pathogens

Article Title: GP73 represses host innate immune response to promote virus replication by facilitating MAVS and TRAF6 degradation

doi: 10.1371/journal.ppat.1006321

Figure Lengend Snippet: ( A ) HEK293 cells (2×10 5 ) were co-transfected with GP73 plasmids at 0, 0.125, 0.25 or 0.5 μg and β-actin plasmid (0.05 μg) along with MAVS or TRAF6 plasmids (0.5 μg) for 24 h. WCLs were subjected to WB with indicated antibodies. ( B ) HEK293 cells (5×10 5 ) were co-transfected with GP73 plasmids at 0, 0.5, 1, 1.5 or 2 μg for 24 h. Endogenous MAVS was determined by WB. ( C ) THP-1-GP73-RNAi cells were collected and endogenous MAVS, TRAF6, IRF3 and IκBα proteins were detected by WB. ( D ) HEK293 cells (5×10 5 ) were transfected with control or GP73 plasmids for 24 h, the mRNA levels of MAVS and TRAF6 were determined by RT-PCR (left panel). THP-1-GP73-RNAi cells were collected to determine the mRNA levels of MAVS and TRAF6 by RT-PCR (right panel). ( E ) Huh7-MAVSR cells were un-infected or infected with HCV (MOI = 2) for 4 days, CHX (cycloheximide, 40 μg/ml) were added for the indicated time points followed by WB analysis. ( F ) HEK293 cells (2×10 5 ) were transfected with GP73 plasmid (1 μg) for 20 h followed by treatment with DMSO, proteasome inhibitor MG132 (20 μM), autophagosome inhibitor 3-MA (1 mg/ml), lysosome inhibitor CQ (Chloroquine, 100 μM) or apoptosis inhibitor Z-FAD-FMK (50 μM) for 4 h. Endogenous MAVS was detected by WB. ( G ) HEK293 cells (2×10 5 ) were co-transfected with GP73 plasmid (0.1 μg) together with TRAF6 plasmid (0.2 μg) for 20 h followed by treatment with DMSO, proteasome inhibitor MG132 (20 μM) or BTZ (Bortezomib, 10 μM), lysosome inhibitor CQ (100 μM) or ER-to-Golgi transport inhibitor BFA (Brefeldin A, 20 μg/ml) for 4 h. WCLs were subjected to immunoblots with the indicated antibodies.

Article Snippet: Antibodies against Flag, HA, c-Myc, TRAF6, Calnexin, Syntaxin 6, p-IκBα and IκBα were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Transfection, Plasmid Preparation, Control, Reverse Transcription Polymerase Chain Reaction, Infection, Western Blot