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Fig. 1. Analysis of organotypic cultures. Organotypic cultures of MDCK-CMV (CMV) (a-d), MDCK-Snail (Snail) (e-h) and MDCK-E47 (E47) cells (i-l) were prepared as described in Materials and Methods and grown for 3 weeks. Histological (a,e,i) and double immunofluorescence analyses for E-cadherin (red) and vimentin (green) (b,f,j) were performed on cryostat sections; nuclei were stained with DAPI (blue). In situ hybridization analysis for Snail (c,g,k) and E47 transcripts (d,h,l) were performed on vibratome sections. Note the infiltration of Snail-expressing cells into the collagen gel and the transdifferentiation to an epithelial non-invading phenotype of the upper layer of MDCK-Snail cultures which re-express E-cadherin and have lost vimentin and Snail expression (arrows in e, f and g). Bars, 50 µm.
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