Figure 3.

Mitophagy was involved in osteoclast-osteomorph recycling

(A) The relative levels of mitochondrial DNA in BMMs, fused osteoclasts, osteoclasts undergoing fission, and re-fused osteoclasts (n = 3).

(B) The ATP production in BMMs, fused osteoclasts, osteoclasts undergoing fission, and re-fused osteoclasts (n = 3).

(C) Western blot analysis of the protein levels of LC3B, ATG5, p62, and GAPDH in BMMs, fused osteoclasts, and osteoclasts undergoing fission (n = 3).

(D) Quantification of LC3B, ATG5, and p62 by immunoblotting.

(E) Representative JC-1 fluorescence images showing fused osteoclasts and osteoclasts undergoing fission (J-aggregate, red; J-monomer, green) (n = 5). Yellow dotted lines indicate osteoclasts. Yellow arrowheads indicate osteoclasts undergoing fission. The J-monomer signal was significantly enhanced in daughter cells, which demonstrated that the mitochondrial membrane potential had decreased.

(F) Representative ROS fluorescence images showing fused osteoclasts and osteoclasts undergoing fission (ROS, green) (n = 5). The red arrowhead indicates osteoclasts undergoing fission.

(G) Western blot analysis of protein levels of TOM20, BNIP3, PINK1, Parkin, and GAPDH in BMMs, fused osteoclasts, and osteoclasts undergoing fission (n = 3).

(H) Quantification of TOM20, BNIP3, PINK1, and Parkin by immunoblotting.

∗p < 0.05 by one-way ANOVA (A, B, D, and H).