Phospho-Akt

The thalidomide analogs CPS49 (30) and CPS11 (31) have been reported to inhibit PI3/AKT signaling in multiple myeloma cells via an anti-angiogenic effect. These compounds are devoid of the teratogenic properties seen with thalidomide and are currently in preclinical development [66]. Compound 30, and to a lesser extent 31, induced a dose-dependent inhibition of proliferation in several multiple myeloma cell lines and reduced phospho-AKT levels [66]. These compounds also inhibited DNA synthesis in cell lines resistant to conventionally used anti-multiple myeloma drugs (e.g. dexamethasone, anthracyclines and melphalan) in a dose-dependent manner. 

Fig. 3. (-)-Linalool does not affect the expression of p-Akt and Akt in the spinal cord of neuropathic mice. (A) Western blot analysis of lumbar spinal cord (L4—L5 segment) 7 days after SNL alone or in combination with linalool (100 mg/kg s.c.) or vehicle treatment Homogenates were from the ipsi(I)-and contra(C)-lateral side of the cord (pool of three animals for each experimental group). (B—D) Densitometric analysis of immunoblots probed with anti-Akt, anti-phospho-Akt (Ser473), and anti-GAPDH antibodies from three different experiments (mean SEM).

PREVENTION OF GLUTAMATE ACCUMULATION AND UPREGULATION OF PHOSPHO-AKT MAY ACCOUNT FOR NEUROPROTECTION AFFORDED BY BERGAMOT ESSENTIAL OIL AGAINST BRAIN INJURY INDUCED BY FOCAL CEREBRAL ISCHEMIA IN RAT... 

Fig. 3. Phosphorylation level of Akt and GSK-3/3 following focal cerebral ischemia. (A) Western blot analysis of phospho-Akt (Ser473) (p-Akt) and total Akt performed on brain cortical homogenates from rats sacrificed 24 h after MCAo shows a trend toward a decrease of p-Akt immunoreactivity in the ipsilateral (I), ischemic, cortex as compared to contralateral (C), nonischemic, side. This trend toward a reduction also occurred for total Akt expression, so that phosphorylation level (expressed as the ratio p-Akt/Akt) is not significantly affected by ischemia. The result is representative of three independent experiments. Histograms in (C) show the results of the densitometric analysis of the autoradioraphic bands corresponding to p-Akt, total Akt, and /3-actin. p-Akt and Akt levels were normalized to the values yielded by /3-actin and Akt phosphorylation was expressed by the ratio of p-Akt/total Akt data are reported as mean S.E.M. ( = 3 per group). The same samples were used for the subsequent western blot analysis of phospho-GSK-3/3 (Ser9) (p-GSK-3/3) and total GSK-3/3 and a representative result of three independent experiments is shown in (B). The results of the densitometric analysis of the bands corresponding to p-GSK-3/3, total GSK-3/3, and a-tubulin are reported in (C). p-GSK-3/3 and total GSK-3/3 were normalized to the values yielded by a-tubulin whereas GSK-3/3 phosphorylation was calculated from the ratio of p-GSK-3/3/total GSK-3/3 data are reported as mean S.E.M. (n = 3 per group).

Liu Q, Afinari L, Chen CS et al (2010) FTY720 shows promising in vitro and in vivo preclinical activity by downmodulating Cyclin D1 and phospho-Akt in mantle cell lymphoma. Clin Cancer Res 16 3182-3192... 

Primary antibodies. Rabbit anti phospho AKT(S473) polyclonal antibody (Cell Signaling Technology), diluted 1 50 in TES-TXIOO. Anti HA polyclonal antibody (Zymed, South San Francisco, CA), diluted 1 100 in TES-TXIOO. 

Remove the blocking solution and place the coverslip cells facing down on 100 pL of the primary antibody solution, either anti Phospho AKT(S473) or anti HA, on the parafilm followed by overnight incubation in the moist chamber at 4°C. 

Celecoxib Does Not Downregulate Akt Phosphorylation Through the Attenuation of PI 3-Kinase Activity. Celecoxib has been shown to reduce phospho-Akt levels in LNCaP cells independent of effects on PI 3-kinase (66), with Akt as a major downstream effector. To determine whether this holds true for the observed downregulation of... 

Given the role of Akt in promoting cell survival under adverse eonditions, it was of interest to investigate the effects of hypoxia on Akt in PC 12 cells (11). We measured a dramatic increase in phospho-Akt (Ser" ) that persisted after 24 hr of exposure to 5% O2. Activation of Akt, and its associated antiapoptotie effects, is typically mediated through PI3K (34,35). Treatment of PC12 cells with wortmannin, a specific inhibitor of PI3K, prior to exposure to hypoxia, eompletely abohshed the effect of hypoxia on phosphorylation of Akt (Fig. 6). Wortmannin did not alter total Akt protein levels (Fig. 6). 

---