Figure 5
STING activation in PDAC CAFs in vitro. (a) Immunoblotting analyses of stimulated CAF1 and Mφ lysate with anti-phospho-STING, anti-STING, anti-phospho-TBK1, anti-TBK1, anti-phospho-IRF3, anti-IRF3,and β-actin antibodies. CAF1 from PDACs and Mφ collected from the abdominal cavity by stimulation with thioglycolate medium were stimulated by DMXAA and harvested after 1 h. (b) The amount of IFN-β in the supernatant was analyzed by ELISA. CAFs and Mφ (6.0 × 105 each) in a 35-mm Petri dish coated with 1000 μL of medium were incubated with DMXAA for 24 h and the supernatants were used(*P < 0.05). Each dot indicates a measure, and the bars indicate the mean. (c) Fold changes of the indicated cytokine and chemokine mRNA levels in DMXAA-treated CAFs relative to the mean of the control. Data are graphed on a log-scale (*P < 0.05). Each dot indicates a measure, and the bars indicate the mean. (d) Schematic of the migration assay. T cells activated by CD3 and CD28 were placed in the upper wells, and supernatants of control CAFs or DMXAA-treated CAFs in the lower wells. After 24 h, migratory cells were lysed and quantified using CyQuant GR fluorescent dye. (e) Migratory cells were measured by fluorometry (*P < 0.05). Each dot indicates a measure, and the bars indicate the mean.
