Effect of GM-CSF stimulation on the expression of PU.1, CIITA, and MHC class II, and on the recruitment of PU.1 to the pIII.

A-H. Quantitative real-time PCR analysis of the mRNA expression of PU.1 (A, E), MHC class II (B, F), total CIITA (C, G), and CIITA driven by pIII (D, H) in mouse splenic pDCs (A-D) or CAL-1 cells (E-H) with (+) or without (-) GM-CSF stimulation. Mouse pDCs and CAL-1 cells were stimulated with 20 ng/ml mGM-CSF for 24h and 10 ng/ml hGM-CSF for 72h, respectively. I. The amount of PU.1 binding to the pIII in GM-CSF-stimulated CAL-1 cells (+) or control cells (-) was determined by a ChIP assay. Binding level of PU.1 (open bar) is expressed as fold change against that of control IgG (closed bar). In E-I, data represent means ± SEMs of three independent experiments performed with duplicate samples. In A-E, the results are expressed as means ± SEMs of triplicate samples, and similar result was obtained in another experiment. *, p < 0.05.