Figure 4.
Moesin depletion boosts bone degradation in both mOCs and hOCs. (A–D) Effect of moesin KO on bone degradation (A and B) and sealing zone (SZ) formation (C and D) in mOCs. (A and B) mOC control (CTL) versus moesin KO (MKO) were cultured for 7 days on bone slices; after cell removal, bone was stained with toluidine blue. (A) Representative images of bone degradation, eroded bone surfaces are delineated by dashed white lines. Scale bar, 50 µm. (B) Quantification of bone eroded surface (%) using semiautomatic quantification. Each circle represents an independent experiment, n = 4, means ± SDs are shown. (C and D) mOC control (CTL) versus moesin KO (MKO) were cultured for 5 days on glass coverslips, detached and then seeded for additional 2 days on bone slices. (C) Representative microscopy images of sealing zones visualized by F-actin staining (phalloidin, white in upper panels and colored-coded intensity in lower panels). Scale bars, 20 and 5 µm. (D) Quantification of the number of sealing zones per bone surface (each circle represents an independent experiment, n = 4, means ± SDs are shown) and of sealing zone thickness (n = 3 independent experiments, 15–20 SZ/condition, 3 locations/SZ). (E–H) Effect of moesin depletion on bone degradation (E and F) and sealing zone formation (G and H) in hOCs. 6 day–differentiated hOCs on glass coverslips treated on day 0 with siCTL or siMoesin (siM) were detached and seeded for additional 24 h on bone slices. (E) Same legend as in A. Scale bar, 100 µm. (F) Same legend as in B (each circle represents a donor, n = 4, means ± SDs are shown). (G) Same legend as in C. (H) Quantification of the number of sealing zones (each circle represents a donor, n = 4) and of sealing zone thickness (n = 3 donors, 15–20 SZ/condition, 3 locations/SZ).

Moesin depletion boosts bone degradation in both mOCs and hOCs. (A–D) Effect of moesin KO on bone degradation (A and B) and sealing zone (SZ) formation (C and D) in mOCs. (A and B) mOC control (CTL) versus moesin KO (MKO) were cultured for 7 days on bone slices; after cell removal, bone was stained with toluidine blue. (A) Representative images of bone degradation, eroded bone surfaces are delineated by dashed white lines. Scale bar, 50 µm. (B) Quantification of bone eroded surface (%) using semiautomatic quantification. Each circle represents an independent experiment, n = 4, means ± SDs are shown. (C and D) mOC control (CTL) versus moesin KO (MKO) were cultured for 5 days on glass coverslips, detached and then seeded for additional 2 days on bone slices. (C) Representative microscopy images of sealing zones visualized by F-actin staining (phalloidin, white in upper panels and colored-coded intensity in lower panels). Scale bars, 20 and 5 µm. (D) Quantification of the number of sealing zones per bone surface (each circle represents an independent experiment, n = 4, means ± SDs are shown) and of sealing zone thickness (n = 3 independent experiments, 15–20 SZ/condition, 3 locations/SZ). (E–H) Effect of moesin depletion on bone degradation (E and F) and sealing zone formation (G and H) in hOCs. 6 day–differentiated hOCs on glass coverslips treated on day 0 with siCTL or siMoesin (siM) were detached and seeded for additional 24 h on bone slices. (E) Same legend as in A. Scale bar, 100 µm. (F) Same legend as in B (each circle represents a donor, n = 4, means ± SDs are shown). (G) Same legend as in C. (H) Quantification of the number of sealing zones (each circle represents a donor, n = 4) and of sealing zone thickness (n = 3 donors, 15–20 SZ/condition, 3 locations/SZ).

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