Figure 3.

A dominant-negative IκBα super-repressor mutant blocks the effect of RIP1 on mTOR. A. U87MG cells were infected with Ad-RIP1+Ad-null or Ad RIP1+ Ad- IκBαM for 24h. B. shows the same experiment in U251MG cells. C. U87 cells were transfected with the putative mTOR promoter linked to luciferase (mTOR-LUC) and co-transfected with FLAG-RIP1 plasmid or empty vector. Introduction of RIP1 results in a significant suppression of mTOR-LUC. Expression of FLAG-RIP1 was confirmed by Western blot with FLAG antibodies. D. this experiment is similar to C except that a mTOR-LUC reporter with mutated NF-κB binding sites was used.