FIGURE 3.

Wortmannin and GS-9820 induce osteoclast retraction. Rat osteoclasts were bathed in HEPES-buffered M199 medium with 15% FBS and antibiotics and imaged using time-lapse phase-contrast microscopy. A–E illustrates 5 different osteoclasts, each shown at 3 times. Images labeled time 0 min illustrate the appearance of cells immediately prior to addition of vehicle, wortmannin (1 μm), PIK75 (1 μm), TGX221 (1 μm), or GS-9820 (1 μm) to the bath. Prior to treatment, osteoclast lamellipodia were well spread and motile. Wortmannin and GS-9820 induced prompt retraction of lamellipodia, which was sustained for at least 30 min. Images are representative of the responses of 9–15 osteoclasts from four to eight independent preparations. Scale bar represents 30 μm for all panels. Images displayed were γ adjusted for clarity of the osteoclast periphery.