Figure 4: Phenotype and PKCα expression in osteoclast cultures transfected with miR-142-3p mimic or PKCα siRNA.

Human CD14+ monocytes were cultured in the presence of M-CSF and sRANKL. (A) Cultures were left untreated or transfected with miR-142-3p mimic, miR-142-3p inhibitor, or control mimic. PKCα expression was assessed by qRT-PCR at day 4, or (B) by western blot at day 6. (C) Cultures were left untreated or transfected with PKCα siRNA, or control siRNA, at day 2. Images of cells stained with Hoescht (nuclei; blue) and LysoTracker (lysosomes, green) taken at day 9. (D) Viability of cultures at day 9 assessed by MTS assay. Expression is relative to untransfected controls. (E) Average number of nuclei per cell of cultures transfected with PKCα siRNA, control siRNA, or untransfected. Data are presented as mean and SD, or representative images, from 3 biological donors. *p < 0.05 relative to untreated control cells. P values were calculated using the Student’s t-test.