Fig. 3.

IRF-8 protein expression was decreased in DCs generated with RM1 cells to the same extent as in DCs transfected with siRNA IRF-8. DCs from murine bone marrow precursors were generated as described in the Fig. 1 legend. a RM1 cells were added in a transwell system and incubated for the first 3 days in cultures. Nuclear extracts were prepared from tumor-treated (DC-RM1) and control (DC-cnt) 7-day-old DCs using the Nuclear Extraction Kit as described in “Materials and methods”. Western blot was performed as described in “Materials and methods”. The results of one representative out of three independent experiments are shown. b 5-day-old DCs were transfected with siRNA for IRF-8 as described in “Materials and methods”. The level of DC transfection using protocol developed by Santa Cruz Biotechnology was between 50 and 60% based on fluorescein conjugated control siRNA expression. Western blot for DCs transfected with IRF-8 siRNA (DC-siRNA) and control siRNA (DC-cnt) was performed as described in “Materials and methods”. The results of one representative out of three independent experiments are shown