mouse monoclonal antibody against human akt1  (Cell Signaling Technology Inc)

Journal: Molecular Medicine Reports

Article Title: MicroRNA-497 inhibits cellular proliferation, migration and invasion of papillary thyroid cancer by directly targeting AKT3

doi: 10.3892/mmr.2017.7345

Figure Lengend Snippet: AKT3 is a direct target of miR-497 in PTC. (A) The binding site for miR-497 in the 3′UTR of AKT3. (B) The relative luciferase activity was determined in HEK293T cells co-transfected with miR-497 mimics or miR-NC, and pGL3-AKT3-3′UTR Wt or pGL3-AKT3-3′UTR Mut. (C) Reverse transcription-quantitative polymerase chain reaction analysis of AKT3 mRNA expression levels. (D) Representative western blot images and quantification of AKT3 and p-AKT3 protein expression levels. Data are expressed as the mean ± standard deviation. *P<0.05 vs. miR-497 NC. PTC, papillary thyroid cancer; miR, microRNA; NC, negative control; Mut, mutant; Wt, wild-type; 3′UTR, 3′ untranslated region; AKT3, RAC-γ serine/threonine-protein kinase; p, phosphorylated.

Article Snippet: Membranes were then incubated with mouse anti-human AKT3 (cat. no. sc-134254; 1:1,000 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (cat. no. sc-32233; 1:1,000 dilution; Santa Cruz Biotechnology, Inc.) monoclonal antibodies, at 4°C overnight.

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Standard Deviation, Negative Control, Mutagenesis

Journal: Molecular Medicine Reports

Article Title: MicroRNA-497 inhibits cellular proliferation, migration and invasion of papillary thyroid cancer by directly targeting AKT3

doi: 10.3892/mmr.2017.7345

Figure Lengend Snippet: AKT3 is upregulated in PTC tissues and negatively correlates with miR-497 expression levels. (A) Reverse transcription-quantitative polymerase chain reaction analysis of AKT3 mRNA expression levels and (B) representative western blot images and quantification of AKT3 protein expression levels in PTC tissues and adjacent normal thyroid tissues. (C) A negative correlation was confirmed between miR-497 and AKT3 mRNA expression in PTC tissues. Data are expressed as the mean ± standard deviation. *P<0.05 vs. normal tissue. PTC, papillary thyroid cancer; miR, microRNA; AKT3, RAC-γ serine/threonine-protein kinase.

Article Snippet: Membranes were then incubated with mouse anti-human AKT3 (cat. no. sc-134254; 1:1,000 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (cat. no. sc-32233; 1:1,000 dilution; Santa Cruz Biotechnology, Inc.) monoclonal antibodies, at 4°C overnight.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: MicroRNA-497 inhibits cellular proliferation, migration and invasion of papillary thyroid cancer by directly targeting AKT3

doi: 10.3892/mmr.2017.7345

Figure Lengend Snippet: Downregulation of AKT3 mimicks roles of miR-497 overexpression on cell proliferation, migration and invasion of papillary thyroid cancer. (A) TPC-1 and HTH83 cells were transfected with AKT3 siRNA or NC siRNA. After transfection for 48 h, western blotting was performed to measure AKT3 protein expression. *P<0.05 vs. NC siRNA. (B) MTT assay analysis of proliferation in TPC-1 and HTH83 cells after transfection with AKT3 siRNA or NC siRNA. *P<0.05 vs. NC siRNA. (C) Cell migration and invasion assays were used to detect migration and invasion abilities of TPC-1 and HTH83 cells after transfection with AKT3 siRNA or NC siRNA. *P<0.05 vs. NC siRNA. (D) TPC-1 and HTH83 cells were transfected with pcDNA3.1-AKT3 or pcDNA3.1. After transfection 48 h, western blot analysis was performed to measure AKT3 protein expression. *P<0.05 vs. pcDNA3.1. Enforced expression of AKT3 partially rescued the suppressive roles of miR-497 on TPC-1 and HTH83 cell (E) proliferation, and (F) migration and invasion. *P<0.05 vs. miR-NC and miR-497 + pcDNA3.1-AKT3. Data are expressed as the mean ± standard deviation. miR, microRNA; AKT3, RAC-γ serine/threonine-protein kinase; NC, negative control; siRNA, small interfering RNA; p, phosphorylated.

Article Snippet: Membranes were then incubated with mouse anti-human AKT3 (cat. no. sc-134254; 1:1,000 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (cat. no. sc-32233; 1:1,000 dilution; Santa Cruz Biotechnology, Inc.) monoclonal antibodies, at 4°C overnight.

Techniques: Over Expression, Migration, Transfection, Western Blot, Expressing, MTT Assay, Standard Deviation, Negative Control, Small Interfering RNA