Journal:

Article Title: Inactivation of Icmt inhibits transformation by oncogenic K-Ras and B-Raf

doi: 10.1172/JCI200418829

Figure Lengend Snippet: Growth factor-stimulated Erk1/2 and Akt1 phosphorylation in Icmt-deficient fibroblasts. (a) Nontransfected Icmtflx/flx and the derivative IcmtΔ/Δ fibroblasts were seeded at equal density and serum-starved overnight as described in the Methods section. Serum-containing medium was then added to the cells. Cells were harvested at the indicated time points and analyzed by immunoblotting with antibodies against phosphorylated Erk1/2 (p-Erk1/2), phosphorylated Akt1 (p-Akt1), and total Erk1/2. (b) Icmtflx/flx and the derivative IcmtΔ/Δ fibroblasts were seeded at equal density and serum-starved overnight. Medium (0.5% serum) supplemented with EGF (50 ng/ml) was added to the cells. Cells were harvested at the indicated time points and analyzed by immunoblotting with antibodies against p-Erk1/2 and total Erk1/2.

Article Snippet: At various times after EGF stimulation, the cells were harvested and total cell extracts were analyzed by immunoblotting with antibodies recognizing phospho-Erk1/2 (phospho-p44/42 MAP kinase E10 mAb; Cell Signaling Technology, Beverly, Massachusetts, USA), phospho-Akt1 [(Ser 473)-R, polyclonal; Santa Cruz Biotechnology Inc.], and total Erk1/2 (p44/42 MAP kinase, polyclonal; Cell Signaling Technology).

Techniques: Western Blot