Journal: Biology of reproduction

Article Title: AKT isoforms 1 and 3 regulate basal and epidermal growth factor-stimulated SGHPL-5 trophoblast cell migration in humans.

doi: 10.1095/biolreprod.112.104778

Figure Lengend Snippet: FIG. 6. Epidermal growth factor-induced phosphorylation of AKT in control shRNAmir, and AKT1, AKT2, or AKT3 shRNAmir cells. Growth factor stimulation, Western blot analyses using antibodies against phosphorylated and total AKT, and densitometrical quantification of signals were performed as described in Materials and Methods. A) Western blots showing expression of p-AKT Ser473, p-AKT Thr308, and total AKT in the different AKT knockdown cell pools. GAPDH was used as a loading control. Representative examples are shown. B) Densitometrical quantification of the different phosphoproteins in EGF-stimulated controls and in AKT1, AKT2, and AKT3 shRNAmir cells. Bar graphs show mean values 6 SD for each of n ¼ 3 different gene-silenced SGHPL-5 cell pools and controls (arbitrarily set at 100%). *P , 0.05; ns indicates not significant compared with control shRNAmir.

Article Snippet: Phosphorylated forms of AKT1 were detected with rabbit anti-human antibodies against p-AKT Ser473 (1:500; Cell Signaling Technology) and p-AKT Thr308 (1:500; Cell Signaling Technology), also detecting the equivalent amino acid residues of AKT2 (Ser474, Thr309) and AKT3 (Ser472, Thr305).

Techniques: Phospho-proteomics, Control, Western Blot, Expressing, Knockdown