Fig. 3

Posttranslational modifications on MIEN1 regulate its function. NIH3T3 cells transfected with the MIEN1 constructs indicated in Fig. 2a, were subjected to scratch wound assays. a–g Confluent monolayers of cells were wounded, and healing of the wound by cell migration was monitored for 18 h. Images were taken at 0 and 18 h. h The fold change in migration was normalized to GFP (empty vector) transfected cells and expressed as the means ± s.e.m of three independent experiments. i–n NIH3T3 cells transfected with indicated GFP-tagged MIEN1 constructs were stained with rhodamine-conjugated phalloidin post wound induction to evaluate effects of ITAM and CAAX motif on filopodia formation. o NIH3T3 cells expressing the GFP vector control and GFP-MIEN1 variants were analyzed for GFP and MIEN1 contents. GAPDH served as a loading control