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. 2013 Sep 17;21(11):2043–2053. doi: 10.1038/mt.2013.187

Figure 1.

Figure 1

Triptolide (TPL) inhibits vesicular stomatitis virus (VSV)-mediated ISG induction in a dose-proportional manner. (a) HEK293 cells were transfected with ISRE-Luc reporter and treated with TPL (2.5, 5.0, and 10.0 nmol/l) or vehicle. Cells were left untreated or were challenged with SeV (40 HAU/ml) or stimulated with interferon (IFN)-α (1,000 U/ml). Luciferase activity was analyzed 24 hours posttransfection and fold activation was determined compared with control. (b) HEK293 cells were treated with 50 nmol/l of TPL and left untreated or were infected with SeV (40 HAU/ml) or stimulated with IFN-α (1,000 U/ml). Western blotting was performed to assess expression of ISG-56 (top panel), RIG-I (middle panel), and β-actin (bottom panel) proteins using ISG56, RIG-I, and β-actin antibodies. (c) Human androgen-independent prostate cancer cell line PC3 cells were preincubated with TPL (12.5, 25.0, and 50.0 nmol/l) or vehicle and infected with VSV-Δ51 (0.005 MOI). The expression of ISGs was assessed using quantitative real-time PCR analysis of total RNA isolated from PC3 cells. (d) Whole-cell extracts (WCEs) were analyzed by immunoblotting with anti-ISG56 (left top panel), anti-PY701 STAT1 (left second panel), anti-ISG15 (left third panel), anti-IRF7 (right top panel), and anti-β-actin (right and left bottom panels) antibodies.