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. 2006 Apr 11;94(9):1300–1310. doi: 10.1038/sj.bjc.6603094

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Copyright © 2006 Cancer Research UK

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Effect of pSUPER.retro infection system on migration and soft-agar colony formation in HT29 cells. The migration assay was performed by using Boyden chambers and 8-μm polycarbonate membranes coated with Matrigel (40-fold dilution). After 24 h of incubation, cells that migrated through the Matrigel-coated membranes were fixed, stained, photographed (A) and counted under light microscopy (B). Regarding the soft-agar colony formation assay, the number of colonies was evaluated 7 days after the seeding in soft agar (C). Control: HT29 wild-type cells; pSUPER(−): HT29 cells infected with vector nonexpressing anti-COX2 shRNA; pSUPER(+): infected HT29 cells expressing shRNA against COX-2 mRNA. In the migration assay samples were tested in the absence and presence of PMA 40 nM. All procedures are described under Material and Methods and data in (B) and (C) represent the mean±s.e.m. of three independent experiments. ^*(P<0.01); #(P<0.05). Bar: 20 μm.