Figure 3
From: Ror2 signaling regulates Golgi structure and transport through IFT20 for tumor invasiveness
IFT20 is required for reorientation of the centrosome toward the direction of cell invasion. (a) Localization of IFT20 at the cis-Golgi in SaOS2 cells. Cells transfected with si-Ctrl or si-IFT20#1 were cultured on FL-gelatin-coated glass coverslips for 6 hr. Cells were then stained with antibodies against IFT20 and cortactin, and counterstained with an antibody, which is conjugated to Alexa Fluor 647 and directed against GM130, a cis-Golgi marker. Boxed regions are magnified on the middle of each image. Note that invadopodia, indicated by a dot-like accumulation of cortactin in the areas of degraded FL-gelatin, are formed in the close vicinity to the Golgi apparatus, where IFT20 is localized. Scale bar, 10 µm. (b) SaOS2 cells transfected with the indicated siRNAs were examined for reorientation of the centrosome using 2D invasion assay (see Methods). The percentages of the edge cells in which the centrosome were detected within the 120° sector emerging from the center of the nucleus and facing toward the space were measured. The red line in the graph indicates the level of expected random orientation of 33%. Data are expressed as mean ± SD of three independent experiments (**P < 0.01, ***P < 0.001, t test).
