Extended Data Fig. 2: Characterization of OCmiR-KD mice.
(a) Representative genotyping image of the Dicerfl/fl mice, Rosa-YFP mice and Dicerfl/fl; Rosa-YFP mice. (b) Representative confocal fluorescent micrographs of YFP expression after Cre-mediated recombination in TRAP+ OCs in the cryosection of subchondral bone from the Dicerfl/fl; Rosa-YFP mice administered with Cre-expressing plasmid encapsulated within the OC-TDS. (c) Gating strategy for the detection of Cre-mediated recombination in OCs in vivo by flow cytometry analysis. The dead cells were excluded by 7-aminoactinomycin D (7-AAD) staining, while the live cells were gated for identifying the OCs with the co-expression of OSCAR (PE) and CTR (APC). The Cre-mediated recombination in OCs was further confirmed by the YFP expression in OCs. (d) Microscopic images of multinucleated osteoclast formation with TRAP staining in BMMs with or without infection of Cre-expressing lentivirus (e) The exosome quantitation (per 106 cells) in cell culture supernatant of OCs differentiated from BMMs with or without infection of Cre-expressing lentivirus. (f-g) Western blot analysis of cellular protein expression of Rab27a and GAPDH (f) and exosomal protein expression of CD63 and CD9 (g). All scale bars: 100 μm. Data are shown as mean ± s.d. All experiments were performed in biologically independent triplicates in each group. The exact p values are shown in figures. Student’s t-test was used for statistical analysis.
