Figure 4

Cell adhesion properties are SETD6 dependent. (A) Cell adhesion to fibronectin. MDA-MB-231 CRISPR SETD6 knock-out (MDA SETD6 KO) cells stably expressing Flag PAK4 wild-type or Flag PAK4 K473R with empty or Flag SETD6 were serum starved over-night. Cells were then plated on a fibronectin pre-coated 96-well plate, followed by crystal violet staining. Crystal violet staining was solubilized and quantified. Error bars are s.d. Statistical analysis was performed for 3 experimental repeats using one-way ANOVA. ***p < 0.001, significance for PAK4 wild-type with Flag SETD6 compared to all other conditions. (B) Actin structures staining. MDA-MB-231 CRISPR SETD6 knock-out (SETD6 KO) cells stably expressing Flag PAK4 wild-type with empty or Flag SETD6, were fixed and stained with phalloidin and DAPI and visualized by a confocal microscope (40×). Scale bar 10 µm. Graph on the bottom shows percent cells with actin structures manually counted. Error bars are s.d. Statistical analysis was performed for n > 49 cells per condition using Student’s t-test. ***p < 0.001.