Fig 6.

Cellular phosphatases are required to inhibit IKKα/IKKβ phosphorylation. (A) i293-ST cells remained uninduced or induced for MCPyV ST expression. Cells were then serum starved before TNF-α was used to stimulate activation of the NF-κB pathway. Cells were lysed directly into SDS sample buffer, and lysates were immunoblotted to detect phosphorylation of IKKα/IKKβ (Ser176/177). GAPDH was used as a loading control, and FLAG blotting detected expression of MCPyV ST. OA, okadaic acid. (B) Densitometry quantification of the Western blots at 5 and 10 min time points was carried out using ImageJ software and is shown as a percentage of relative densitometry normalized to the loading control GAPDH. Standard deviations of 3 replicated experiments are shown.