Figure 3-6923.

The effect of overexpression of EYFP-p62WT or EYFP-p62ΔUBA on osteoclast resorptive activity. EYFP-p62WT, EYFP-p62ΔUBA, or EYFP stably expressing RAW264.7 cell lines were cultured on bovine bone slices in the presence of RANKL (100 ng/ml). A: After 14 days, cells were fixed and stained for either TRACP (a–c) or rhodamine-phalloidin (F-actin) (d–f) and visualized by light microscopy and confocal microscopy, respectively. Cells were then removed, and resorptive lacunae were assessed by scanning electron microscopy (g–i). F-Actin rings are marked with arrows. Quantitative analysis shows the mean nuclei numbers (B) and sizes (C) of osteoclasts, percentage of osteoclasts forming F-actin rings on bovine bone slice (D), and percentage of bovine bone slice surface occupied by resorption lacunae (E). *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars = 50 μm.