Antifoam

VS-5584 and ABT-737 were obtained from Selleck (Shanghai, China). Both compounds were dissolved in DMSO and stored at -20° for in vitro studies. For in vivo studies, both agents were dissolved in the SX-1292 oral vehicle [1% sodium carboxymethyl cellulose, 0.5% sodium lauryl sulfate (SLS), and 0.05% antifoam; Eli Lilly, Shanghai, China] and administered by gastric lavage [13 (link),14 (link)]. Antibodies of p-AKT (Ser 473, #9271), p-AKT (Thr 308, #9275), p-S6 ribosomal protein (S6, Ser 235/236, #2211), S6 (#2317), p-p70 S6 kinase 1 (S6K1, Thr 398, #9209), S6K1 (#2708), Bcl-2 (#2870), Bcl-xL (#2762) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118) were obtained from Cell Signaling Technology (Danvers, MA). The concentration of primary antibodies utilized in this study was 1: 1,000, except for p-S6 (1: 10,000) and GAPDH (1: 10,000). The caspase inhibitors z-VAD-fmk and z-DVED-fmk were obtained from Calbiochem (Shanghai, China).

Icaritin (purity 98%) was purchased from Shenogen Pharma Group (Beijing, China). Icaritin was dissolved in dimethyl sulfoxide (DMSO) for in vitro studies. The final concentration of DMSO in the culture media was maintained at 0.1%, which showed no cytotoxicity to HCC cells. For in vivo studies, icaritin was dissolved in the SX-1292 oral vehicle [1% sodium carboxymethyl cellulose, 0.5% sodium lauryl sulfate (SLS), and 0.05% antifoam; Eli Lilly, Shanghai, China] and administered by gastric lavage. D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), FTY720 and SP600125 were obtained from Sigma (St. Louis, MO); SKI-II (4-[[4-(4-Chlorophenyl)-2-thiazolyl]amino]phenol) and sorafenib were purchased from Tocris Bioscience (Ellisville, Mo). Sphingosine 1-phosphate (S1P) and C6 ceramide were obtained from Avanti Polar Lipids, Inc. (Alabaster, AL). All antibodies utilized in this study were purchased from Cell Signaling Technology (Beverly, MA).