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. 2012 Oct 31;287(51):43019–43029. doi: 10.1074/jbc.M112.388694

FIGURE 4.

FIGURE 4.

p65 (21–186) fragment inhibits the NF-κB signaling required for RPS3 nuclear translocation via competing RPS3 off p65. A, HEK293T cells were transiently transfected with either EGFP vehicle control or EGFP-tagged p65 (21–186) truncated mutant, followed by stimulation with TNF (50 ng/ml) for indicated period. Whole cell lysates were derived, separated, and immunoblotted directly for serine 209 phosphorylated RPS3 (p-RPS3), with β-actin as a loading control. B, whole cell lysates (Input) from HEK293T cells, transiently transfected with either EGFP or EGFP-tagged p65 (21–186) truncated mutant and stimulated as indicated, were immunoblotted directly or after IP with RPS3 antibody for RPS3 and importin-α (Imp-α). C and D, whole cell lysates (Input) from HEK293T cells, transiently transfected with either EGFP vehicle control or EGFP-tagged p65 (21–186) truncated mutant, were immunoblotted directly or after IP with RPS3 antibody for GFP fusion proteins with GFP antibody (C) or for endogenous p65 with p65 (C-20) antibody (D).