(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.
Fig. 2. TRAF6 induces translocation of PH-Akt-GFP to cell membranes and de novo protrusions. (A) A PH-Akt-GFP expression vector reporter was co-transfected with expression vectors encoding a second activator protein, as indicated above each panel, into either HEK293 (a-c) or RAW264.7 (d-f) cells for 24 hours. Transfection of only the activator protein, followed by fixation and phalloidin-TRITC staining in order to visualize polymerized actin (g-i). (B) PH-Akt-GFP expression vector reporter co-transfected with a TRAF6 expression vector into HEK293 cells in either the absence or presence of 25 µM of Ly294002 (LY) and 10 mM methyl-ß-cyclodextrin (MbCD) inhibitors for the indicated treatment time (a-i). Two different sets of cells are shown for MbCD. In one (d-f), a long exposure time reveals the details of the filopodial network. In another (g-i), a shorter exposure reveals that the PH-Akt-GFP is relocalized from cell membranes to the cytoplasm, following treatment with MbCD. A control set shows the co-transfected vectors evaluated in the presence of the DMSO vehicle for an equivalent treatment time (j-l) with a long exposure time to allow visualization of filopodia.
