Supplementary Fig. S3.

(A–B). PAK1 is required for complete activation of NF-κB downstream of IκB in HCEC-1CT. (A) The effect of PAK1 on p65 was evaluated using siPAK1 or scrRNA. Cells were treated with TNFα and analyzed for PAK1, p65, p-IκB-α, and IκBα expression in nuclear and cytoplasmic cell fractions by WB. α-Tubulin and Fibrillarin were utilized as loading controls. siPAK1 impeded nuclear accumulation of p65 on TNFα treatment independent of IκB degradation. (B) NF-kB luciferase assay of TNFα treated cells with or without IPA-3 pretreatment. IPA-3 blocked TNFα induction of NF-kB.