Journal: Oncotarget

Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

doi: 10.18632/oncotarget.11337

Figure Lengend Snippet: ( A ) Changes in morphology and gene expression in MZF-1 60– 72 construct-transfected stably cloned MB-231 cells. β-actin was used as a control, and c-Myc was used as a marker for transfected cells. ( B ) Confocal microscopy showing the distribution of the Elk-1 and MZF-1 proteins. Cells were stained with antibodies against Elk-1 and MZF-1, followed by the appropriate FITC- or rhodamine-conjugated secondary antibodies. Confocal slices of 0.5 and 0.6 μm thicknesses were obtained, and images were obtained focusing the center of the nucleus. “N” denotes the nucleus; “C” denotes the cytosol. ( C ) ChIP assays indicating the binding activity of endogenous Elk-1 and MZF-1 to the PKCα promoter in various cells. The assays were performed using an MZF-1 (left panel) or Elk-1 (right panel) antibody. ( D ) Visualization and quantification of cell migration and proliferation of modified MB-231 cells. ( E ) Tumor growth in nude mice after xenografts of modified MB-231 and Hs578T cells (left panels). Tumors removed from the mice were weighed (middle images and graph) and sliced before histological examination (right images). ** p < 0.01 compared with the empty vector-transfected MB-231 group, ## p < 0.01, compared with the empty vector-transfected Hs578T group.

Article Snippet: The cells were then fixed in 4% paraformaldehyde for 30 min and permeabilized with 0.1% saponin, 2% goat serum (Vector Laboratories) and 0.02% NaN3 for 15 min. Elk-1 and MZF-1 were visualized using their particular antibodies (1:200) followed by FITC-conjugated goat anti-mouse IgG antibody and rhodamine-conjugated goat anti-rabbit IgG antibody (Rockland Immunochemicals, Inc., Gilbertsville, PA.), respectively.

Techniques: Gene Expression, Construct, Transfection, Stable Transfection, Clone Assay, Control, Marker, Confocal Microscopy, Staining, Binding Assay, Activity Assay, Migration, Modification, Plasmid Preparation