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Fig. 8. LPR1 is required for maintenance of membrane protrusions in ovarian epithelial cancer cells. (A) Proteins from SK-OV-3 cells that were either untransfected (control), or transfected with pEGFP alone or in combination with the control double-stranded RNA (RNAic) or two double-stranded siRNAs designed to target LPR1 (RNAi1 and RNAi2) were analyzed by western blotting for LPR1 expression. Immunoreactive species were quantified by scanning densitometry and pixel densities (normalized to control cells) are shown below each lane of the gel. (B) SK-OV-3 cells were co-transfected with pEGFP in combination with either RNAic or RNai2, and actin organization was visualized by fluorescence microscopy. Stars denote cells expressing the EGFP marker. Bars, 20 µm. (C) SK-OV-3 cells were transfected with EGFP alone or in combination with either RNAic or RNAi2. Phalloidin-stained filopodia were counted only in EGFP-expressing cells from the three different categories, and the averages of each were plotted. Error bars denote s.d. The average number of filopodia per cell in EGFP expressing cells was 56.7 (s.d. 20.6, n=21). EGFP-expressing cells co-transfected with RNAic had an average of 57.5 filopodia per cell (s.d. 13.5, n=26). EGFP expressing cells co-transfected with RNAi2 had an average number of 27.3 filopodia per cell (s.d. 12.3, n=25).
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