FACS, MTT, and colony-formation assays of NIH3T3-flagOVA66 and NIH3T3-mock cells.
A. FACS analysis of cell cycle of the two transfected NIH3T3 cells. OVA66 overexpression in NIH3T3-flagOVA66 cells significantly promote cell cycling compared with NIH3T3-mock cells. B. OVA66 overexpression promoted proliferation of NIH3T3-flagOVA66 cells in vitro. Proliferation of NIH3T3 and NIH3T3 transfected with pFlag-OVA66 and pFlag-vector were analyzed by MTT assay for 7 days. C. Colony formation assay in 6-well plates. NIH3T3-flagOVA66 and NIH3T3-mock cells were seeded at 0.4×103 cells per well. After 2 weeks of cultivation, cells were fixed with 4% paraformaldehyde and stained with crystal violet. Numbers of colonies formed by NIH3T3-flagOVA66 and NIH3T3-mock were calculated. The data are represented as mean ± SEM (n = 3), *P≤0.05, **P≤0.01.
