FIG 2.

IpaJ is sufficient for blocking NF-κB translocation to the nucleus. (A, D) HeLa cells were treated with TNF-α for 30 min or remained untreated, after which they were fixed and stained with anti-p65 (green) and 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B, E) HeLa cells transfected with plasmids expressing HA tag, HA-IpaJ, or HA-IpaJ (C90A) were treated with TNF-α for 30 min or remained untreated, after which they were fixed and stained with anti-p65 (green), anti-HA (red), and DAPI (blue). (C, F) HeLa cells were infected with wild-type (WT), a pSPI12 mutant, or ΔpSPI12-pipaJ. After 3 h, cells were washed and treated with TNF-α and gentamicin. At 30 min post-TNF-α induction, they were fixed and stained with anti-p65 (green) and DAPI (blue). The measurement of p65 in HeLa cells were performed in duplicate, and the percentage of area in nucleus occupied by p65 was analyzed to demonstrate the translocation ability of p65 into the nucleus (D, E, F). ***, P ≤ 0.001.