IFN-γ inhibits T-antigen expression in glial cells.

<p>A. PHFA cells were transfected with pCDNA 3.1 plasmid encoding T-antigen. Cells were treated with IL2 (6 ng/ml), Rantes (10 ng/ml), IFN-γ (100 ng/ml), IL-1β (5 ng/ml), IL13 (5 ng/ml), and MCP1 (6 ng/ml) for 48hr. Whole cell protein lysates were processed for the detection of T-antigen, phospho-Stat1 and total-Stat1. The same membranes were stripped off and probed for anti-tubulin as loading control. In lane 1, whole cell protein lysates from cells transfected with pcDNA3.1 vector DNA were also processed as controls of transfection. B. Bar graph representation of normalized T-antigen expression signal from panel A. C. Stat1 signaling is induced by soluble immune mediators secreted by activated immune cells. PHFA cells were transfected with an expression plasmid encoding T-antigen and treated with conditioned media from either uninduced or induced PBMCs. Western blot analysis of protein extracts was performed to assess phosphorylated and total protein levels Stat1 and Stat3. Tubulin was probed as loading control in the same membranes. D. Normalized expression levels of phospho-Stat1, total-Stat1, phospho-Stat3, and total-Stat3 from the Western blot studies in panel C were presented as bar graph.