FIGURE 5.

Effect of Cr-TRP16 on the expression and subcellular localization of NF-κB. A, effects of Cr-TRP16 expression on NF-κB p50, p65, and IκBα expression. HeLa cells were transfected with Cr-TRP16 as described under “Experimental Procedures.” Whole cell lysates were subjected to Western blot analysis. The FLAG-tagged Cr-TRP16 was detected with anti-FLAG antibody. B and C, HeLa were transfected with the plasmids Cr-TR16 or C15S Cr-TRP16 for 30 h and then stimulated with TNF-α (1 nm) for 6 h. Nuclear and cytoplasmic protein extracts were prepared using nuclear extraction kit (Active Motif) according to the manufacturer's instructions. Protein concentration was determined with Bradford reagent. The nuclear and cytoplasmic extracts were subjected to Western blot analysis. The level of actin served as a loading control. NE, nuclear extract; CE, cytoplasmic extract; WCE, whole cell extract.