Mk 2206

Eribulin was kindly provided by Eisai Inc. (Tokyo, Japan) and Sapporo Medical University for in vitro and in vivo use, respectively. MK-2206 was obtained from ChemScene (Monmouth, NJ, USA). Pazopanib was obtained from AdooQ BioScience (Manassas, VA, USA). Stock solutions of these reagents were generated by dissolving the powder in 100% dimethyl sulfoxide (DMSO; Sigma–Aldrich St. Louis, MO, USA) at 10 mM. HT1080, SK-LMS-1 and SW872 cell lines were purchased from ATCC (Manassas, VA, USA). Both cell lines were maintained in DMEM (Sigma–Aldrich) containing 10% fetal bovine serum, 1% penicillin–streptomycin and 2 μM L-glutamine.

The Akt-inhibitor, MK-2206 (Chemscene, Monmouth Junction, NJ), was used. The hNPCs were cultured in micromass in chondrogenic basal medium supplemented with TD-198946 (10 nM) and MK-2206 (10 nM to 1 μM) for seven days for alcian blue staining and real-time PCR. The hNPCs were also cultured with DMEM supplemented with TD-198946 (10 nM) and MK-2206 (100 nM) (TD + MK2206) for 60 min for western blot analysis.