Figure 3
From: SUMOylation of Grb2 enhances the ERK activity by increasing its binding with Sos1
SUMOylation of Grb2 at K56promotes cellular transformation. (A) Comparable expression levels of Grb2WT and Grb2K56R in stable NIH/3T3 cell lines generated by polyclonal lentiviral infections were determined by Western blotting. (B) Stable NIH/3T3 cell lines were seeded in 2 ml of medium containing 5% FBS with 0.35% agar at 5 × 104 cells/well and layered onto the base. The photographs were taken 3 weeks later. (C) Vasculogenic mimicry assay for stable NIH/3T3 cell lines. Each cell line at a density of 1 × 104 was plated in each well which was coated with matrigel and photographs were taken with microscope 20 hours later. (D) Stable NIH/3T3 cell lines at 60% confluence were stimulated with 5 μg/ml of Doxycycline (Doxy) for 24 or 36 hours. The ERK activities were determined by Western blotting. (E) Densitometric readings were obtained with ImageJ V1.45 software, and the signals of phosphorylated Erk were normalized to that of total Erk. Data are representative of three independent experiments.