Pan akt inhibitor gsk690693

Matrigel matrix was obtained from Corning. Insulin, human recombinant, zinc solution was purchased from Thermo Fisher Scientific. TGF-β1 and BMP-9 were obtained from R&D Systems. Inhibitors of ALK1 (LDN193189), PI3K inhibitor (LY294002), Src family inhibitor (PP2), and MEK inhibitor (PD98059) were obtained from Sigma–Aldrich. Pan-Akt inhibitor (GSK690693) was purchased from Selleckchem. The following antibodies were all purchased from Cell Signaling: P-Smad1/5/9, Smad1, P-Akt serine 473, P-p42/44 MAPK, Src, and IR. β-actin was purchased from Sigma–Aldrich, anti-HA was from Roche, and Endoglin (P3D1), Endoglin (A-8), ALK1, and EPDR1 antibodies were purchased from Santa Cruz Biotechnology.

Immortalized normal liver cell LO2 and human HCC cell lines including MSK-Hep1, SNU-475, SNU-423, HepG2, Huh7, Huh1, SNU-182, and Hep3B were purchased from the ATCC and cultured in DMEM high glucose (HyClone) supplemented with 10% fetal bovine serum (FBS), and the cells were maintained at 37 °C in 5% CO₂ incubator. Cisplatin (DDP) was purchased from Selleck, and 30 μM was used to treat cells. Pan-Akt inhibitor GSK690693 was purchased from Selleck, and 10 nM was used to treat cells.