Forced expressions of hRasGRP4 and its splice variants in transcription-translation assay and in mammalian cells.A. Cell-free transcription translation assay. The vectors encoding full-length hRasGRP4, its splice variant 5 and 6 were subjected to cell-free transcription translation assay using rabbit reticulocyte lysate. Resulting products were subjected to immunoblotting using anti-hRasGRP4- horseradish peroxidase (HRP). B. Expression of hRasGRP4 and its splice variants in mammalian cells. The vectors encoding full-length hRasGRP4, its splice variant 5 or 6 were transfected into HEK293 cells. Cell lysates were subjected to immunoblotting using anti-hRasGRP4- HRP, anti-phospho or anti-pan P44/42 mitogen-activated protein kinase (MAPK) (Erk1/2). Representative results of three independent experiments are shown.
Hashimoto et al. Arthritis Research & Therapy 2011 13:R154 doi:10.1186/ar3470