PMC

Flow cytometry in osteosarcoma cells. SaOS-2 and LM8 cells were treated with Veh (A & C) or 10 μM 2-ME (B & D) for 24hrs and analyzed using a FACS Scan unit.

Ligand specific cell cycle arrest in osteosarcoma cells. MG63 cells were treated with Veh, vehicle (A), 10 μM of 2-ME (B), E2, 17β-estradiol (C) and 16α-OHE, 16α-hydroxyestradiol (D) for 24 hr and analyzed using a FACS Scan unit.

Dominant negative PKR blocks 2-ME-mediated cell cycle arrest. MG63 cells were transfected with the plasmid pCDNA3-PKR-K296R (A & B) or empty vector pcDNA neo (C &D) and treated with Veh or 10 μM 2-ME for 24 hr. The cells were harvested and analyzed by flow cytometry.

2-ME treatment increases p21 protein levels. Cytoplasmic extracts prepared from osteosarcoma cells treated with Veh and 10 μM 2-ME for 24 hrs were analyzed by Western blot hybridization using anti-p21 (A), anti-p53 (C) (Cell Signaling Technology) anti-p27 (B) (Santa Cruz Biotechnology) and anti-actin (D) (Sigma) antibodies.

2-ME induces cell death in SaoS-2 (A) and LM8 (B) osteosarcoma cell lines. Osteosarcoma cells were treated with Veh or 5 μM of 2-ME for 72 hrs. The cells were harvested and the proliferation was determined by MTS assay. Values are the mean ± SE (N=3 replicate cultures). The absence of error bar denotes a line thickness greater than the error. *P≤ 0.05 versus Veh.