Journal: PLoS ONE

Article Title: Mechanisms of GnRH-Induced Extracellular Signal-Regulated Kinase Nuclear Localization

doi: 10.1371/journal.pone.0040077

Figure Lengend Snippet: (A) HeLa cells were seeded in 12 well plates, transduced with Ad mGnRHR and kept in reduced (0.1%) serum for 16 hours prior to stimulation for varied periods with 100 nM GnRH or 1 µM PDBu. Whole cell protein extracts were then immunoblotted for phospho Ser217/221 MEK1/2 (ppMEK1/2), phospho Thr183/Tyr185 ERK (ppERK) and for total ERK (ERK1 and/or 2) as indicated. (B) HeLa cells were seeded in 96-well imaging plates, transduced with Ad mGnRHR and kept in reduced (0.1%) serum for 16 hours prior to stimulation for 5 min with 1 or 100 nM GnRH as indicated. Control cells (Ctrl.) were treated with medium alone. The cells were then fixed and stained for endogenous ppERK, ERK and DAPI before image acquisition and analysis (as described in the ). The figure shows representative images and the lower images show “segmented” ERK staining, with lines indicating the perimeters of the nuclei and cells obtained using automated image analysis algorithms, as described in the . Each of the image panels corresponds to a width of approximately 250 µm and represents approximately 1/20 th of the area captured in each field of view.

Article Snippet: Ser217/221 phosphorylated MEK1/2 were detected using a rabbit anti-ppMEK1/2 monoclonal antibody (clone 41G9, 1∶1000, Cell Signaling Technology).

Techniques: Transduction, Imaging, Staining

Journal: eLife

Article Title: Registered report: Kinase-dead BRAF and oncogenic RAS cooperate to drive tumor progression through CRAF

doi: 10.7554/eLife.11999

Figure Lengend Snippet:

Article Snippet: Rabbit α ppMEK1/2 , Antibody , Cell Signaling Technology , 9121 , , .

Techniques: Cell Culture, Transfection, Bradford Assay, Detection Assay, Western Blot, Staining, Transduction