Journal:

Article Title: Growth Factor-Independent Proliferation of Erythroid Cells Infected with Friend Spleen Focus-Forming Virus Is Protein Kinase C Dependent but Does Not Require Ras-GTP

doi:

Figure Lengend Snippet: Expression of dominant-negative Ras in uninfected and SFFV-infected HCD-57 cells transfected with H-rasN17. Cell lysates were prepared for Western blot analysis from uninfected and SFFVP-infected HCD-57 cells cotransfected with a vector expressing the dominant-negative H-rasN17 mutant (lanes 2, 4, and 6) or a control vector (lanes 1, 3, and 5) plus the pEGFP-N2 N-terminal fusion vector expressing GFP and the neomycin resistance gene. Cells expressing GFP were purified by FACS and evaluated for H-rasN17 expression 2 days after transfection (lanes 1 to 4) or 3 months after G418 selection (lanes 5 and 6) using an anti-Ras antibody that detects both endogenous and mutant forms of the Ras protein. Protein loading was determined by immunoblotting with an anti-Raf-1 antibody.

Article Snippet: For Raf-1 and MAPK immune complex kinase assays, protein was purified from 1 mg of lysate using anti-Raf-1 or Erk-2 polyclonal (C14) antibodies from Santa Cruz Biotechnology.

Techniques: Expressing, Dominant Negative Mutation, Infection, Transfection, Western Blot, Plasmid Preparation, Mutagenesis, Purification, Selection

Journal:

Article Title: Growth Factor-Independent Proliferation of Erythroid Cells Infected with Friend Spleen Focus-Forming Virus Is Protein Kinase C Dependent but Does Not Require Ras-GTP

doi:

Figure Lengend Snippet: PKC is required for activation of MAPK but not Raf-1 or MEK in uninfected and SFFV-infected HCD-57 cells. Uninfected and SFFV-infected HCD-57 cells were treated with the PKC inhibitor staurosporine (Stauro) (40 nM) or bisindolylmaleimide (Bis) (20 μM) or the PKA inhibitor HA1004 (40 μM). Uninfected HCD-57 cells were then stimulated with Epo for 15 min, while SFFV-infected HCD-57 cells were left unstimulated. Cell lysates were assayed in immune complex assays for MAP kinase activity (A), Raf-1 kinase activity (B), and MEK kinase activity (C). Protein loading was determined by immunoblotting with the appropriate antibody. MEK−, GST-kinase-inactive MEK.

Article Snippet: For Raf-1 and MAPK immune complex kinase assays, protein was purified from 1 mg of lysate using anti-Raf-1 or Erk-2 polyclonal (C14) antibodies from Santa Cruz Biotechnology.

Techniques: Activation Assay, Infection, Activity Assay, Western Blot

Journal: Molecular cell

Article Title: The Functional Proximal Proteome of Oncogenic Ras Includes mTORC2

doi: 10.1016/j.molcel.2018.12.001

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RAF1 Human Recombinant Protein , Origene , Cat# TP301983.

Techniques: Western Blot, Transduction, In Situ, Recombinant, Lysis, Protease Inhibitor, Staining, Magnetic Beads, Clone Assay, Bicinchoninic Acid Protein Assay, Isolation, Labeling, Viability Assay, RNA Sequencing Assay, Sequencing, Expressing, CRISPR, Mass Spectrometry, Mutagenesis, shRNA, Plasmid Preparation, Luciferase, Software, Blocking Assay