Fig. 5
UFM1 suppresses the metastatic potential and epithelial-to-mesenchymal transition of gastric cancer in PDK1-dependent manner. a UFM1 associates with PDK1 in gastric cancer. Immunoprecipitation using PDK1 antibody was performed in AGS cell lysates (up panel). Stable AGS cells (down panel) were collected, lysed, and cell lysates were applied to immunoprecipitation with UFM1 antibody. b UFM1 promoted PDK1 ubiquitination. 293 T cells were cotransfected with constructs as indicated. PDK1 was immunoprecipitated with an anti-PDK1 antibody, and the ubiquitinated PDK1 was visualized by Western blot analysis using an anti-Ub antibody. c Immunofluorescence images showing the changes in PDK1 in stable AGS cells. d Stable AGS cells were treated with Control siRNA or PDK1 siRNA then cell lysates were applied in western blot analysis. e The stimulatory effect of UFM1 downregulation on AGS cell migration and invasion was rescued by PDK1 siRNA transfection; scale bar, 50 μm. f Quantitative results of (e) is show. The data are presented as the mean ± SD (*P < 0.05; **P < 0.01; ns, no significance)