Fig. 5

Immunodetection of K18 and β-actin protein expression in KGN cells mock transfected (siCTL) or transfected with 100 nM siRNA to KRT8 and KRT18. a Representative in-cell western dual detection of K18 protein (green), β-actin protein (red), and combined (merged image-yellow) following exposure to either Lipofectamine™ or equimolar concentrations of a non-targeting siRNA (siCTL) as controls, or 100 nM siRNA to KRT8/18 (siKRT8/18). Diminished expression of K18 in the single and merged images was evident following 72 h exposure to siKRT8/18; b Quantification of K18/β-actin expression compared to siCTL following siKRT8/18 transfection. The mean (± SEM) following treatment for three independent, replicate experiments is depicted. c Representative image of immunofluorescent staining of K18 and actin filaments in cultured KGN cells. Keratin (K18) filament expression stained with FITC (green); β-actin filament expression stained with Phalloidin (red); nuclei stained with DAPI (blue); and merged image of FITC, Phalloidin and DAPI. Bar graph represents quantification of K18 fluorescence (mean relative light units, RLU ± SEM). (n = 6 independent images; different letters denote differences, P < 0.05)