FIG. 7.

Net and Net(L16A) exhibit different transcriptional responses to JNKK and Ras. (A) NIH 3T3 cells were cotransfected with the reporter (1 μg of Palx8-TK-LUC; all lanes), the empty vector lacking Net coding sequences (lanes 2 to 5), or expression vectors for Net [250 ng of pTL2Net; lanes 6, 8, 10, 12, 14, 16, 18, 20, and 22], Net(L16A) [250 ng of pTL2Net(L16A); lanes 7, 9, 11, 13, 15, 17, 19, 21, and 23], Ha-Ras (pRasCTBx2; 100 ng [lanes 2, 8, and 9], 250 ng [lanes 3, 10, and 11], 500 ng [lanes 4, 12, and 13], 1,000 ng [lanes 5, 14, and 15]), or JNKK (pSRα-mJNKK; 100 ng [lanes 16 and 17], 250 ng [lanes 18 and 19], 500 ng [lanes 20 and 21], 1,000 ng [lanes 22 and 23]). Fold activation was calculated relative to the basal activity of the reporter in the presence of empty vector (pSG5). (B) Expression levels of Net and Net(L16A) were assessed by Western blotting using an antibody against Net (PAb 375). The specific band is indicated on the left.