PDGF-AA-induced dorsal ruffle formation is diminished in WIP ^−/− fibroblasts.

<p><b>a</b> Control (WIP^+/+) and WIP^−/− primary murine fibroblasts were serum starved over night (0 min) or serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and FITC-secondary antibody and imaged in a Zeiss microscope to identify dorsal ruffles (white arrow). Magnifications of the boxed areas are shown as right panels. <b>b</b> The percentage of WIP^+/+(black) and WIP^−/− (white) cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. <b>c</b> WIP^−/− primary fibroblasts were lentivirally transduced to express control cherry or WIP-cherry, starved and incubated with PDGF-AA for 8 or 15 min. Fixed cells were imaged. <b>d</b> The percentage of WIP^−/− cells expressing cherry (white) or WIP-cherry (black) and forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. <b>e</b> Representative western blot of PDGFRα expression in soluble lysates of lung-derived fibroblasts from WIP^+/+ and WIP^−/− mice. Numbers indicate relative expression levels of the protein to control fibroblasts determined by densitometry. ß-actin labeling confirmed equivalent protein loading control. <b>f</b> WIP^+/+ (black) and WIP^−/− (white) primary fibroblasts were grown in the presence of serum (left panel) and stained with anti-PDGFRα plus labeled secondary antibody and analysed by FACS. The mean fluorescence intensity of positive cells in both populations is represented. Right panel, WIP^+/+ (black) and WIP^−/− (white) primary fibroblasts were starved (time 0) and stimulated with PDGF-AA and stained as above. Percent of mean fluorescence intensity relative to starvation is represented. Arbitrary units (a.u.). * p<0,05.