Figure 1

IL-27-mediated activation of STAT1 and STAT3. (A) A549 cells were treated with IL-27 (50 ng/mL) for up to 72 hours. The tyrosine phosphorylated, or activated, forms of STAT1 and STAT3 (P-STAT1 and P-STAT3) as well as the total amounts of the transcriptional factors (T-STAT1 and T-STAT3) were detected by Western blot. (B) Seven human NSCLC cell lines (H1703, H292, H157, H1437, H460, H1650, and H358) were cultured with the diluent of IL-27 (0.1% PBS/BSA) or IL-27 (50 ng/mL) for 24 hours and the activated and total amounts of STAT1 and STAT3 proteins were measured by Western blot. The densitometric measurements of total amounts of STAT1 and STAT3 were taken using Image J1.45o. The values above the figures represent relative density of the bands normalized to GAPDH. (C-D) A549 cells were treated with IL-27 (50 ng/mL) for 15 minutes, and stained with anti-tyrosine phosphorylated STAT1 (C) (green) and STAT3 (D) (green) antibodies for immunofluorescence microscopy (50 × magnification). The cells were counterstained with DAPI (blue). The white arrows indicate cells with nuclear activation of STAT1 or STAT3 by IL-27 treatment. Scale bar, 100 μm. (E) Expression of IL-27 receptor (TCCR) on cultured A549 cells. (F) Expression of IL-27 receptor (TCCR) on A549 cells after treatment with or without IL-27 (50 ng/mL) for 24 hours.