FIG 5.

Increased osteoclastogenesis in RANKL-primed precursors infected with S. aureus is only partially mediated by TLR2 and TLR9. (A–C) BMDMs were cultured with 35 ng/mL RANKL + CMG 14-12 supernatant for 2 days. The cells were then infected with S. aureus at an MOI of 25 in the absence of RANKL, using gentamicin protection. For all, *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. If not denoted with asterisks, the differences between genotypes were not statistically significant. Error bars denote the SD, with n = 3 technical replicates per group. The data are representative of 3 independent experiments. (A) Representative images of TRAP-stained cells at day 2 postinfection. (B) After 2 days of infection, the cells were fixed and stained, and TRAP⁺ multinucleated (3 ≥ nuclei) osteoclasts were quantified. A two-way ANOVA was used to assess the effects of genotype and treatment, and Sidak’s test was used to compare each vehicle to S. aureus for each genotype. (C) The counts were normalized to the vehicle and were analyzed using a one-way ANOVA. Dunnett’s multiple-comparisons test was used to compare each genotype to the WT.