FIG 3.

The ITAM-containing adaptors Dap12 and FcRγ do not participate in B. burgdorferi phagocytosis and signaling. (A) BMDMs from wild-type (WT) and Dap12- or FcRγ-deficient mice were stimulated with B. burgdorferi at an MOI of 10 for 6 h. Supernatants were collected, and the concentrations of secreted IL-6 and TNF-α were determined by an ELISA. Data are shown as means and standard errors of the means of results from three independent experiments. *, P < 0.05. (B) BMDMs from wild-type and Dap12- or FcRγ-deficient mice were stimulated with Bb-GFP for 60 min at 37°C. Any inhibitors or blocking agents used were added for 30 min prior to Bb-GFP stimulation. Following Bb-GFP stimulation, cells were fixed in 1% paraformaldehyde and stained with 2% goat serum and 1% saponin. Noninternalized B. burgdorferi bacteria were stained with anti-B. burgdorferi antibodies followed by a secondary anti-rabbit Alexa Fluor 594 antibody. Phagocytosis is shown as percent internalized bacteria per cell. *, P < 0.05.