siRNA Silencing of HBx and Dll4 Genes

The siRNA sequences specific for HBx and Dll4 and scrambled siRNA were designed and purchased from Silencer® Select Pre-Designed and Validated siRNA (Life Technologies, USA). Stealth si-HBx RNA sequences have three duplexes as follows: siHBx-199 5’-AGG TGA AGC GAA GTG CAC ATT-3’ [16 (link)], siHBx-260 5’-GAA TGT TGC CCA AGG TCT TAC ATA A-3’ and siHBx-371 5’-GGG AGG AGA TTA GAT TAA AGG TCTT-3’. Stealth si-Dll4 RNA sequences have three duplexes as follows: HSS182569 5’-CCT CTC CAA CTG CCC TTC AAT TTCA-3’, HSS123069 5’-GCC TAT CTG TCT TTC GGG CTG TCAT-3’ and HSS123063 5’-ACC TCC ATT TGT GAT TAG ACA TGTT-3’. Transfection of siRNA was carried out using Lipofectamine RNAiMax Reagent (Life Technologies, USA). After 24–72 hr of transfection, cells were collected for further analysis.