Figure 1.

JSI-124 effects on ImC and DC differentiation in vitro. (A) Gr-1⁺ ImCs were isolated from spleens of CT26 tumor-bearing mice. DCs were generated from bone marrow progenitors using GM-CSF and IL-4 in the presence of CT26 CM as described in details previously (24) and CD11c⁺ DCs were isolated using magnetic beads. DCs and ImCs were treated with 0.5 μM JSI-124 or vehicle control (DMSO, vc) for 24 hrs in the presence of CT26 CM. Cells were then collected and Western blotting with antibodies against different members of STAT family was performed. (B) Gr-1⁺ ImCs were isolated from spleens of tumor-bearing mice, then treated with 0.5 μM JSI-124 or vc and cultured for 7 days in the presence of GM-CSF and CT26 CM. Cell phenotype was evaluated by flow cytometry. Cumulative results from three performed experiments are shown. (C) Four millions Gr-1⁺ ImCs were isolated from spleens of C3 tumor-bearing C57BL/6 (CD45.2⁺) mice and transferred into congenic C3 tumor-bearing B6.SJL-PtrcaPep3b/BoyJ (CD45.1⁺) mice (tumor size 1 cm in diameter). Mice were then treated with JSI-124 or DMSO for 3 days. On day 4 mice were sacrificed and phenotype of donor's CD45.2⁺ splenocytes was evaluated by flow cytometry. Each group included 4 mice.