Fig. 2.

Spontaneous DC activation in Vps34^f/f;CD11c-Cre mice. (A) Splenocytes, lymph node, and lung cells were prepared from mice, stained with anti-CD11c and -CD11b antibodies and with anti-CD40, -CD80, -CD86, -K^b, -D^b, or isotype control antibodies, and were analyzed by flow cytometry. Representative plots from at least two experiments with six mice per group are shown. (B) Splenic DCs were purified by FACS and cultured in complete medium either alone or in the presence of the indicated stimuli for 24 h. Culture supernatants were collected to measure IL-6, TNFα, and IL-10 by cytometric bead array (CBA). A representative of three experiments is shown. The error bars indicate the mean ± SD of triplicate wells. (C) BMDCs were activated as in B, and culture supernatants were collected at 48 h for measurement of IL-6 and TNFα by CBA. A representative of three experiments is shown. The error bars indicate the mean ± SD of triplicate wells. (D) BMDCs (10⁴ cells) were activated with the indicated numbers of heat-killed L. monocytogenes in a 24-well plate, and 48 h later culture supernatants were tested for IL-1β secretion by DCs. A representative of three experiments is shown. The error bars indicate the mean ± SD of triplicate wells.