Journal:

Article Title: Enterolactone Inhibits Insulin-Like Growth Factor-1 Receptor Signaling in Human Prostatic Carcinoma PC-3 Cells 1 2

doi: 10.3945/jn.108.101832

Figure Lengend Snippet: Enterolactone inhibits IGF-1-induced tyrosine phosphorylation of IGF-1R in a dose- (A,B) and time-related (C,D) manner. (A) PC-3 cells were starved in serum-free medium for 24 h and pretreated with indicated doses of enterolactone for 0.5 h followed by stimulation with IGF-1 for 15 min. After treatment, cellular proteins were isolated and immunoprecipitation and immunoblotting were performed. (B) Quantified data of A. The intensity of phospho IGF-1R and total IGF-1R protein signals obtained in A was quantified using Chem Doc densitometry software (Quantity One, version 4.5.0; Bio-Rad). The densitometry data were normalized to IGF-1R levels and those of the control (lane 1). (C) PC-3 cells were starved for 24 h, followed by treatment with enterolactone (60 μmol/L) for 0.5 h, then IGF-1 was added, cells were incubated and harvested at various time points, and levels of phospho-IGF-1R and IGF-1R were determined. (D) Quantified data of C. Values are means ± SEM, n = 4. Means without a common letter differ, P < 0.05. ENL, Enterolactone.

Article Snippet: The intensity of phospho IGF-1R and total IGF-1R protein signals obtained in A was quantified using Chem Doc densitometry software (Quantity One, version 4.5.0; Bio-Rad).

Techniques: Phospho-proteomics, Isolation, Immunoprecipitation, Western Blot, Software, Control, Incubation

Journal:

Article Title: Enterolactone Inhibits Insulin-Like Growth Factor-1 Receptor Signaling in Human Prostatic Carcinoma PC-3 Cells 1 2

doi: 10.3945/jn.108.101832

Figure Lengend Snippet: Enterolactone inhibits phosphorylation of AKT (A,C), its downstream targets (B,D,E,G), and phosphorylation of ERK1/2 (F,H). PC-3 cells were starved in serum-free medium for 24 h. Then, cells were pretreated with indicated doses of enterolactone for 0.5 h. IGF-1 was added to stimulate cells for 15 min (A,B,F) and 6 h (E). After treatment, cellular proteins were isolated and immunoblotting was performed. (C,D,G,H) Quantified data of immunoblotting. The densitometry data of phospho-AKT, phospho-p70S6K1, phospho-GSK-3β, cyclin D1, and p-ERK1/2 obtained in A,B,E, and F were normalized to GAPDH levels and those of the control (lane 1). Values are means ± SEM, n = 4. Means without a common letter differ, P < 0.05. ENL, Enterolactone.

Article Snippet: The intensity of phospho IGF-1R and total IGF-1R protein signals obtained in A was quantified using Chem Doc densitometry software (Quantity One, version 4.5.0; Bio-Rad).

Techniques: Phospho-proteomics, Isolation, Western Blot, Control