Figure 5

Cell proliferation and migration assays. (A) MS1 cells in 96-well plate were transfected with 1 pmol synthetic siRNAs. 48 h later, transfection medium was replaced with regular medium containing BrdU labeling reagent. Incubation lasted for 16 h before proliferation assay was performed. *denotes p < 0.05 (B) EOMA cells in 96-well plate were transfected with 1 pmol synthetic siRNAs. 48 h later, BrdU labeling solution was added into medium. Incubation lasted for 16 h before proliferation assay was performed. (C) Cell migration assay. EOMA cells in 6-well plate were transfected with 25 pmol synthetic siRNA. 48 h later, cells were placed onto trans-well membrane to allow migrate for 2 h. Cells were counted under microscope. (D) Representative spheroids with sprouting vessels. MS1 cells in 6-well plate were transfected with 25 pmol synthetic siRNA. 24 h later, cells were suspended in hanging drops to allow cellular aggregation. Spheroids were embedded in collagen gel to culture for 7 days. (E) Quantification of sprouts growing from the spheroids. 30 spheroids were examined for each transfection. *denotes p < 0.05.