Figure 4
From: Septins are critical regulators of osteoclastic bone resorption
Trench-making human OC: Confocal IF detection of SEPT9 and clathrin show that they strongly co-localize at the sites of the OC engaged in active bone resorption and to those located within the boundaries of the trench. (A) 3D z-projection of trench resorbing OC. Scale bar: 20 µm. B) 3D z-projection using phase contrast to visualize the perimeters of the trench made by the OC shown in A. (C) Cross-section (xz) through the length axis of the OC and trench as indicated by a white dashed line in A. Scale bar: 10 µm. (D–F) A single confocal plane (0.55 µm) (xy) showing an enlargement of the pseudopod area indicated by a white box in A. within the cavity. Scale bar: 10 µm. (G) Line scan through the area marked by a white dashed line in D. (H and I) A single confocal plane (0.55 µm) (xy) at the bone surface level. Scale bar: 20 µm. (J) An enlargement of the area indicated by the white box 1 in H. Scale bar: 5 µm. (K) Line scan through the area marked by a white dashed line in J. (L and M) A single confocal plane (0.55 µm) (xy) at the trench level within the cavity. Scale bar: 20 µm. (N) An enlargement of the area indicated by the white box 2 in L. Scale bar: 5 µm. (O) Line scan through the area marked by a white dashed line in N. Colors: green, phalloidin/F-actin; red, IF staining of SEPT9; blue, IF staining of clathrin; grey, phase contrast; purple, co-localization of SEPT9 and clathrin. Representative of 5 experiments.
