The conditional knock-down of SSX inhibits the proliferation, survival and cell cycle progression of the melanoma cell line DFW in vitro.

<p>A) Graphic representation of the shRNA sequence (complementary to SSX1 to SSX9) ligated into shRNA vectors for stable and doxycycline regulated shRNA expression (see material and methods). B) Western blot showing SSX expression in control-shRNA and SSX-shRNA transfected cells 24 hours after doxycycline addition to the culture medium. C) Cell colony quantification in control and SSX-shRNA transfected DFW cells grown in the presence of doxycycline for 8 days. D) Cell proliferation curves determined by counting the number of alive cells in control and SSX silence cultures using trypan blue staining. E) S-phase cell cycle progression determined by BrdU incorporation in a fluorescence activated cell sorter (FACS). F) Percentage of cells at G1, S and G2 phases of the cell cycle in control and SSX shRNA knocked down DFW cells over 96 hours period.