Akt (Cell Signalling Technology (CST), 9272S, Leiden, Netherlands), Phospho-Akt (Ser473) antibody (CST 4060S, Leiden, Netherlands), [beta]-Actin Antibody (CST 4967, Leiden, Netherlands) antibodies were used for western blotting.
With the increase of LY294002 concentration, phosphorylation levels of PI3K and
AKT were notably reduced compared with the untreated cells (P<0.05, P<0.01 or P< 0.001, Figure 4A).
It's main players are protein kinase B (
Akt) and the mechanistic target of rapamycin (mTOR).
However, no variations were observed in the total
Akt protein level.
Study author Charles Hoeffer said, "
AKT is a central protein that has been implicated in a bevy of neurological diseases yet we know amazingly little about it".
We found that cotreated 8-Br-cGMP with hypoxia showed a significant decrease in protein levels of PI3K p110[alpha], PI3K p110[beta], and phosphorylated
AKT in comparison with the hypoxia group.
The mathematical model is constructed by extending the
AKT phosphorylation model in [21].
Measurement of
Akt, p-Akt, GSK-3[beta], and p-GSK-3[beta] by Western Blot.
In fact, mTORC2 phosphorylates protein kinase C (PKC)[delta], PKC[zeta], PCK[gamma], and PkC[epsilon] that are involved in cytoskeleton assembly and cell migration, besides
AKT on serine 473 [18-21] (Figure 2(c)).
On the other hand, considering the known close relationship between
AKT and SREBP2, the ARRB-dependent signaling pathway may be included in cholesterol regulation.
For dual staining of
Akt and NF-[kappa]B, the sections were stained with a rabbit polyclonal antibody against rat
Akt (1 : 300, Abcam, USA), a rabbit monoclonal antibody against rat NF-[kappa]B p50 (1: 100, Abcam, USA), an Alexa Fluor 594 goat anti-rabbit IgG (1: 200 dilution, Abcam, USA), and an Alexa Fluor 488 goat anti-rabbit (1:200 dilution, Abcam, USA).
In patients with type 2 diabetes undergoing a resistance exercise program, there are increases in PI3K activity,
Akt content, GLUT4, and GS activity.