Characterization of Dectin-1 conditional knock-out macrophages.

(A) The Dectin-1 mRNA level of Dectin-1 control (Cre 0) and Dectin-1 knockout (Cre Tg) peritoneal macrophages was quantified by quantitative real-time RT-PCR. Values are representative of data obtained from three mice. (B) The surface protein level of Dectin-1 was measured by flow cytometry on the Dectin-1 control (Cre 0) and Dectin-1 knockout (Cre Tg) peritoneal macrophages. Plots are representative of data obtained from six mice. (C) The protein level of several receptors (CD11b, Mannose Receptor MR, TLR4, TLR2, SIGNR1, CD36) and markers (F4/80 and CD14) on the macrophage surface was determined by flow cytometry on the Dectin-1 control (Cre 0) and Dectin-1 knockout (Cre Tg) peritoneal macrophages. (D and E) Released [3H]arachidonic acid is expressed as the percentage of [3H]arachidonic acid in the culture medium divided by the total incorporated [3H]arachidonic acid in murine peritoneal macrophages. The release of [3H]arachidonic acid was determined after incubation for 120 min of peritoneal Dectin-1-control and Dectin-1-knockout macrophages with non-opsonized C.albicans (ratio 1∶3) or PMA (100 nM). The data represent the means±SE of three separate experiments. (F) TNFα production by Dectin-1 control (Cre 0) and Dectin-1 knockout (Cre Tg) macrophages after 24 h of stimulation with heat-killed C.albicans (ratio 1∶3), ZNO (2 µg/mL) or LPS (100 ng/mL). Data are the means±SE of three separate experiments. ** (p<0.01) and * (p<0.05) indicates a significant difference compared with the respective Cre 0 or Cre Tg control. § (p<0.05) indicates a significant difference between Cre 0 and Cre Tg.