FIG. 5.

Conditional deletion of the extreme C terminus of mTOR in ES cells. (A) The 3′ terminus of the mTOR gene, the targeting vector, and the correctly targeted mutant locus are shown. Solid triangles in the latter two sequences indicate loxP sites. Asterisks indicate translational termination codons of mTOR. The positions of restriction enzyme sites (B, BamH; H, HindIII) and probes for Southern blotting and the locations of PCR primers (arrowheads) are also shown. (B) Southern blot and PCR analyses. Solid arrowheads indicate bands corresponding to the wild-type locus, whereas open arrowheads indicate bands corresponding to the targeted locus. WT, wild-type ES cells; Flox/+, heterozygous targeted ES cells; Flox/Flox, homozygous targeted ES cells. (C) Northern blot analysis. (D) Western blot analyses showing the protein levels of mTOR, 4E-BP1, and CDK4.