Plasmids
The human AKT (NM_001014431.1) coding region with GFP-Tag was cloned
using PCR and was ligated into the pEGFP-N1 vector. The origin PCR
primers for AKT cds sequence were as follows: Sense primer: 5’ CCG GAA
TTC ATG AGC GAC GTG GCT ATT 3’; Anti-sense primer: 5’CGC GGA TCC CCG GCC
GTG CCG CTG GCC GA 3’. The constitutively active form of Akt (CA-Akt,
myrAkt delta4-129) was purchased from Addgene. Flag-Ubiquitin was a kind
gift from Professor Changan Jiang (Sichuan University, China). The
plasmids were introduced into cells using Lipofectamine 3000 according
to the manufacturer’s protocol.