Figure 5.
Moesin role in bone degradation is independent of its role in osteoclast fusion. (A) Experimental design of moesin depletion in late stages of hOC differentiation. (B) Effect of moesin depletion (siMoesin) on hOC fusion: quantification of fusion index after microscopy analysis of 10 day-differentiated hOCs on glass coverslips treated on day 6 with siCTL or siMoesin. Each circle represents a single donor, n = 7, SDs are shown. (C) Western blot analysis of moesin and P-ERM expression levels after moesin depletion in late stages of hOC differentiation. Representative western blot analysis (left) and quantification of moesin and P-ERM signals (right), normalized to actin. Predicted molecular weight are indicated. Each circle represents a single donor, n = 8, SDs are shown. (D–I) Effect of moesin depletion (siMoesin) in mature hOCs on bone degradation (D), morphology of the resorbed area (E), and sealing zone (SZ) formation (F–I). 10 day-differentiated hOCs on glass coverslips treated on day 6 with siCTL or siMoesin were detached and seeded for additional 24 h on bone slices. (D) Representative images of bone degradation (left, scale bar, 50 µm) and quantification of bone eroded surface (%) using semi-automatic quantification (right). Each circle represents a single donor, n = 5. (E) Quantification of the percentage of trenches (n = 2 independent experiments, SDs are shown). (F and G) (F) Representative microscopy images of sealing zone visualized by F-actin staining (phalloidin, white, scale bars, 20 µm); and (G) quantification of the number of sealing zones (number of SZ per bone surface (left) and the percentage of area covered by SZ (right). Each circle represents a single donor, n = 6. (H and I) Effect of moesin depletion (siMoesin) in mature hOCs on SZ organization (H) thickness (I). (H) Representative microscopy images of sealing zones visualized by F-actin and vinculin staining (phalloidin in pink and vinculin in green). Scale bars, 10 µm. (I) Quantification of sealing zone thickness (n = 3 donors, 15 SZ/condition, 2 locations/SZ). n.s., not significant. Source data are available for this figure: SourceData F5.

Moesin role in bone degradation is independent of its role in osteoclast fusion. (A) Experimental design of moesin depletion in late stages of hOC differentiation. (B) Effect of moesin depletion (siMoesin) on hOC fusion: quantification of fusion index after microscopy analysis of 10 day-differentiated hOCs on glass coverslips treated on day 6 with siCTL or siMoesin. Each circle represents a single donor, n = 7, SDs are shown. (C) Western blot analysis of moesin and P-ERM expression levels after moesin depletion in late stages of hOC differentiation. Representative western blot analysis (left) and quantification of moesin and P-ERM signals (right), normalized to actin. Predicted molecular weight are indicated. Each circle represents a single donor, n = 8, SDs are shown. (D–I) Effect of moesin depletion (siMoesin) in mature hOCs on bone degradation (D), morphology of the resorbed area (E), and sealing zone (SZ) formation (F–I). 10 day-differentiated hOCs on glass coverslips treated on day 6 with siCTL or siMoesin were detached and seeded for additional 24 h on bone slices. (D) Representative images of bone degradation (left, scale bar, 50 µm) and quantification of bone eroded surface (%) using semi-automatic quantification (right). Each circle represents a single donor, n = 5. (E) Quantification of the percentage of trenches (n = 2 independent experiments, SDs are shown). (F and G) (F) Representative microscopy images of sealing zone visualized by F-actin staining (phalloidin, white, scale bars, 20 µm); and (G) quantification of the number of sealing zones (number of SZ per bone surface (left) and the percentage of area covered by SZ (right). Each circle represents a single donor, n = 6. (H and I) Effect of moesin depletion (siMoesin) in mature hOCs on SZ organization (H) thickness (I). (H) Representative microscopy images of sealing zones visualized by F-actin and vinculin staining (phalloidin in pink and vinculin in green). Scale bars, 10 µm. (I) Quantification of sealing zone thickness (n = 3 donors, 15 SZ/condition, 2 locations/SZ). n.s., not significant. Source data are available for this figure: SourceData F5.

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