Fig. 2.

AZA-mediated elimination of the tumor cells promotes differentiation of tumor-derived CD11b cells into CD11c⁺ MHC class II-positive antigen-presenting cells. CT-26 tumors were harvested from syngeneic tumor-bearing mice on days 12–14 after tumor cell inoculation. Whole tumor single cell suspension has been prepared as described in “Materials and methods”. Tumor-infiltrated CD11b cells were purified by positive selection using magnetic beads. Fresh-isolated CD11b cells were washed with PBS, enumerated, placed in six-well plates (2 × 10⁶ per well) in complete medium and cultured in the presence of AZA (5 μM) and GM-CSF (20 ng/ml). On day 7, cells were collected, washed with PBS, stained with antibody against CD11b, F4/80, CD11c, I-A^d, CD86 and CD45, analyzed by flow cytometry. Results of one representative experiment out of three are shown