Figure 4: SENP7S dictates the nuclear localization of Axin.
From: Novel SUMO-Protease SENP7S Regulates β-catenin Signaling and Mammary Epithelial Cell Transformation
(A) To identify Axin1, proteins from cytoplasmic (C), cell membrane (M), and nuclear (N) fractions from MCF10-2A cells treated with siSENP7S and siNT (control) were subject to western blot analysis. CBX5, EGFR, and GAPDH served as a loading control for the nuclear, membrane, and cytoplasmic fraction, respectively. (B) MCF10-2A cells were seeded on to coverslips and transiently transfected with either empty vector (EV) or SUMO3, or co-transfected with SUMO3 in the presence of wild-type SENP7S or mutant SENP7 (SENP7m). After 24 hrs, cells were stained with an Axin1 antibody (red) and DAPI (blue). Images were acquired by fluorescence microscopy using a 20X objective. Insets represent enlarged cells. (C) Cells from immunofluorescence images (B) were counted and categorized based on their Axin1 cytosolic (C) and nuclear (N) localization and the number of cells were presented as percentages. C < N staining was not detected in any group. Data shown as mean ± SEM of two independent experiments. One-way ANOVA was used to compare between groups and p < 0.05 was considered significant.
