Figure 3.
GW 654652 inhibits cell motility. MM.1S cells were pretreated with GW654652 (10 μg/mL) or with DMSO control (approximately 6 hours) and then plated on fibronectin-coated tissue-culture plates (35 × 10-mm plates; BD PharMingen), in the presence or absence of VEGF (100 ng/mL). Dynamic changes in MM cell morphology that mediate migration were assessed by phase-contrast TLVM. (A) Decreased cell motility after GW654652 treatment. Photographs were acquired at 2-minute intervals starting 3 hours after plating. Images shown are at 10-minute intervals, and arrows identify the movement of a single cell. Dashed rectangles identify those cells enlarged in panel B. Apostrophes indicate minutes. (B) Decreased ruffling/formation of lamellipodia and uropods after GW654652 treatment. Closed arrows indicate uropod formation, and open arrows indicate lamellipodia formation. (C-D) Cell perimeter (C) and surface area (D) illustrate morphologic changes on GW654652 treatment. Time-lapse micrographs were analyzed using SlideBook 4.0 software.