FIG. 6.

Normal myeloid cell functions in the absence of Cybr. (A) BMDCs were cultured from wild-type and Cybr-deficient mice and analyzed for the ability to upregulate MHC class II and CD86 by flow cytometry after LPS stimulation. Dashed (leftmost) line, isotype control; solid line, wild type; bold line, Cybr knockout. (B) BMDCs were also tested for their ability to produce cytokines in response to LPS stimulation in vitro. Error bars indicate standard errors of the mean. WT, wild type; KO, knockout. (C) Using Alexa Fluor-labeled dextran or whole bacteria, Cybr-deficient antigen presenting cells were tested for macropinocytosis or phagocytosis. BMDCs were incubated with Alexa dextran at 4 or 37°C for 30 min before being washed and analyzed by flow cytometry (upper panel). Alternatively, activated peritoneal macrophages were incubated with Alexa staphylococci for 30 min, washed, and analyzed by flow cytometry for cell-associated bacteria (bottom panel). Wild-type or Cybr^−/− mice were injected intraperitoneally with 1 ml of 3% thioglycolate solution, and peritoneal exudate cells were recovered 24 h later. (D) Cells were quantified by counting on a hemacytometer. Error bars indicate standard errors of the mean. WT, wild type; KO, knockout.