FIGURE 5.

miR-29 knockdown supports RAW264.7 cell commitment to the macrophage lineage. GFP and GFP_Sponge RAW264.7 cells were induced toward osteoclastic differentiation by 24-h RANKL treatment (30 ng/ml) in the presence or absence of DOX (500 ng/ml). A, lineage commitment was assessed by phagocytosis assay. The cells were incubated for 1 h with pHrodo bacterial particles (250 μg/ml). Cells that have phagocytosed the particles display red fluorescence (arrowheads). Nuclei were stained using DAPI reagent. Representative images of the cultures were captured using 10× magnification. Scale bar, 100 μm. B, phagocytosis was quantified by measuring rhodamine fluorescence and normalized to DAPI fluorescence (n = 6). C and D, macrophage marker mRNAs (C) and osteoclast marker mRNAs (D) were quantified by qRT-PCR and normalized to Hprt mRNA (n = 3). *, significantly different from no DOX, p < 0.05.