Fig 6.

Net1A expression is required for efficient cell spreading. (A) MCF7 cells were transfected with control or Net1A-specific siRNAs. Three days later the cells were serum starved, trypsinized, and replated on collagen-coated coverslips. At different times, the cells were fixed and stained for F-actin (red) and DNA (blue). Shown are results of a representative experiment. (B) Quantification of cell area in siRNA-transfected cells. MCF7 cells were transfected with the control of siRNAs specific for Net1, Net1A, or the dual isoform (Net1/Net1A). The cell area after replating was measured using ImageJ software. Shown are the averages of three independent experiments. Error bars are standard errors of the means. *, P < 0.05; **, P < 0.01; ***, P < 0.001. The inset shows a representative Western blot for Net1 isoform-specific knockdown. (C) Quantification of cell shape. Cells were transfected with the siRNAs shown and replated on collagen-coated coverslips for 60 min. Cell shape (length/width) was measured. Shown are the averages of three independent experiments. Bars are median values. *, P < 0.05.