FIGURE 7.

Cordycepin sensitivity of reporter genes requires 3′ sequences. (A) NFκB responsive firefly luciferase reporter genes with terminator or 3′ UTR sequences from the CXCL1 (pCXCL1-Ter, pCXCL1-3′Ter), histone H4 (pHistH4), and β globin (pHBB) genes were constructed. (Black circles) NFκB binding sites; (arrow) promoter start; (gray boxes) SV40 derived 5′ and 3′ UTR sequences; (white box) luciferase coding region; (black boxes) 3′ sequences from the indicated human genes. Sizes of these regions are not to scale. (B) TNF-induced chemokine mRNA induction in A549 cells is also sensitive to cordycepin. mRNA levels were determined as previously after 1 h TNF induction as described for Figure 3A. Error bars represent 1 standard deviation over three qPCR replicates. This is a representative experiment of several performed under similar but not identical conditions. (C) These reporters were transfected into A549 cells and the cells were treated with TNF in the absence or presence of cordycepin for 2 h. The induction in the control cells (with TNF, no cordycepin) was set at 1 for each construct. Error bars in this figure are standard deviations over three independent biological replicates. Statistical significance in a paired T-test for comparison with noncordycepin treated is indicated: (*) P ≤ 0.05, (**) P ≤ 0.01.