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. 2009 Jul 20;119(8):2399–2411. doi: 10.1172/JCI37155

Figure 5. RIG-I and MDA-5 induce expression of the proapoptotic BH3-only members of the Bcl-2 family Noxa, Puma, Bim, and Bik.

Figure 5

(A) 1205Lu cells were treated with pppRNA or poly(I:C) (3 ng/ml) for 24 hours, and levels of Noxa, Puma, Bim, Bad, Bik, Bid, and Hrk mRNA were measured by quantitative RT-PCR. Relative mRNA levels compared with mock-transfected cells are depicted. Mean ± SD of 3 independent experiments is shown. *P ≤ 0.05 and NS compared with mock-transfected cells. (B) The melanoma cell lines 1205Lu, WM239A, WM278, and WM793 were treated with pppRNA or poly(I:C) (5 ng/ml), and Noxa and Puma proteins were quantified by immunoblotting. β-Actin served as loading control. Blots are representative of 2 independent experiments. (C) Left: Expression of the BH3-only proteins Noxa, Puma, Bim, Bad, Bik, and Bid was inhibited by treatment with respective siRNAs for 48 hours. Thereafter, cells were treated with pppRNA (left), poly(I:C) (5 ng/ml; right), or transfection reagent alone and analyzed for apoptotic cells by FACS. Annexin V–positive and propidium iodide–negative cells are represented. Mean ± SD of 3 independent experiments is shown. *P ≤ 0.05 compared with control siRNA–transfected cells treated with pppRNA.