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Figure 7


Fig. 7. Keratin organisation in periplakin siRNA transfected wound edge cells. Transiently transfected cells that had reached confluency were wounded 72 hours after transfection. (A) K8 immunofluorescence in control transfected cells. (B) K8 immunofluorescence in periplakin siRNA transfected cells. (C) Fluorescence intensity at wound edge compared with monolayer (adjusted to 100%) in control and periplakin siRNA transfected cells. Fluorescence intensity was measured in Image J programme from raw Zeiss LSM 510 images (Merged Z-stacks) from 20 cells in three independent wounds at wound edge and in epithelial monolayer 5 to 7 cell rows away from the wound edge. (D) Cell survival after siRNA transfections. Cell survival was measured using CellTiter cell proliferation kit (Promega). Mean and standard deviation (absorbance units at 490 nm) of three measurements are shown. (E) Phase contrast microscopic images of scratch wound closure of control and Periplakin siRNA transfected MCF-7 monolayers. (F) Quantification of the wound closure in MCF-7 and HeLa cells. The width of the wounds at 20 hours time point were measured from photomicrographs and displayed as percentage of remaining wound width from the start of the assay. Mean and s.e.m. from three independent transfections are shown.





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