Figure 2: LSF-induced inhibition of human melanoma cell proliferation in vitro. (a) Net proliferation of SK-Mel28 cells transiently transfected with the indicated amounts of each vector was quantified by crystal violet staining two days after transfection. Induction of LSF-overexpression was confirmed by immunoblotting. Arrow and arrowhead indicate endogenous and exogenous (DsRed fused) LSF, respectively. (b) FACS-based cell cycle analysis of SKMel28 cells transfected with 3 μg DsRed-LSF expression vector or empty vector three days before analysis. (c) Immunoblot and immunofluorescence detection of LSF in SK-Mel28 cell clones that stably over-express the DsRed- LSF fusion protein or empty vector. (d) Proliferation curve of each cell clone by counting the cell number. Data represent mean cell number/well ± SD of three independent experiments. **P<0.01.