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. 2023 Mar 2;222(4):e202206121. doi: 10.1083/jcb.202206121

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© 2023 Zhu et al.

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Figure 3. — Dual Mmp9/Mmp14 deletion does not affect Cthrc1 levels or galectin-3 expression in osteoclasts. (A) Cthrc1 (green) immunofluorescence stained with an anti-Cthrc1 polyclonal antibody (ab85739; Abcam) and F-actin (red) imaged following phalloidin staining of wild-type and DKO osteoclasts. Results are representative of three independent experiments. Scale bar, 20 μm. (B) Relative expression levels of glycosylation-related transcripts retrieved from DAVID GO categories were assessed in DKO osteoclasts as compared with wild-type osteoclasts (normalized to 1; n = 3 biological replicates). (C) Schematics of galectin-3 monomer domains are presented. (D) Galectin-3 (green) stained with an anti–galectin-3 monoclonal antibody (#125401; Biolegend; clone M3/38) and Mmp9/Mmp14 (cyan) immunofluorescence (upper and lower panels, respectively) of a femur section from a Csf1r-Cre^+/−/Rosa^Tomato/+ mouse. BM cavity, bone marrow cavity. Scale bar, 30 μm. Results are representative of three independent experiments. (E) The relative mRNA levels of Lgals3 in DKO osteoclasts were assessed by qPCR compared to wild-type osteoclasts (normalized to 1). Data are presented as mean ± SEM (n = 3 biological replicates). Statistical significance was assessed using unpaired two-sided Student’s t test.