Article Title: AKT3 promotes prostate cancer proliferation cells through regulation of Akt, B-Raf TSC1/TSC2
Figure Lengend Snippet: AKT3 overexpression and siRNA knockdown affected expression of signaling proteins in PC-3 cells Protein expression of total AKT, AKT3, phospho-AKT S473, phospho-AKT T308, c-Myc, Skp2, p21 Cip , p27 Kip , cyclin D1, cyclin E, GSK3α, phospho-GSK-3α S21, GSK3β, phospho-GSK-3β S9, B-Raf, TSC1, TSC2, mTOR, phosphor-mTOR S2448, p70S6K, phopsho-p70S6K T421/S424 in PC-3 vector control, PC-3 overexpressing AKT3, PC-3 scramble control, and PC-3 AKT3 siRNA knockdown was assayed by Western blotting. Protein abundance of α-tubulin and β-actin were used as loading control.
Article Snippet: Protein expression was determined by Western blotting using following antibodies: phospho-AKT S473, phospho-AKT T308, β-catenin, GSK3α, GSK3β, phospho-GSK-3α S21, phospho-GSK-3β S9, cyclin D1, cyclin E, mTOR, and total AKT antibodies were from Cell Signaling (Danvers, MA, U.S.A.); c-Myc, TSC1, TSC2, B-Raf, and phospho-p70S6K T421/S424 antibodies were from Epitomics (Burlingame, CA, U.S.A.); p21Cip1 , p27Kip1 , and Skp2 antibodies were from Santa Cruz (Santa Cruz, CA, U.S.A.); AKT3, phospho-mTOR S2448 and p70S6K antibodies were purchased from Millipore (Billerica, MA, U.S.A.); β-actin antibody was from Novus (Littleton, CO, U.S.A.);α-tubulin antibody was from Sigma (St. Louis, MO, U.S.A.).
Techniques: Over Expression, Expressing, Plasmid Preparation, Western Blot