Figure 5. IN04 treatment disrupts the cytoskeleton arrangement and F-actin ring formation of osteoclasts. Osteoclast precursors derived from C57BL/6J mice were seeded on bone slices and differentiated in the presence of DMSO or INO4 for 6–9 days. Mature osteoclasts were stained with Alexa Fluor 488-conjugated phalloidin and DAPI, and the actin ring formation and sealing zone were visualized by confocal microscopy. (A) Representative images of IN04- and DMSO-treated osteoclasts. (B) Horizontal cross sections of selected osteoclasts. Two lines in the middle of the cell represent positions of horizontal and vertical cuts, respectively. Rectangles are orthogonal views of X- and Y-sections, respectively. Arrows in red indicate sealing zones (SZ). (C) 3D rendering of a lateral view. A DMSO-treated osteoclast was invaded in a resorptive pit indicated by an arrow in green while an IN04-treated cell was on the surface of the bone slice.