Figure 5.

Induction of mesenchymal features by primary human skin keratinocytes under cytokine treatments. Primary keratinocytes were cultured in growth medium to near confluence. A: Plasticity of skin keratinocytes in mesenchymal morphology was evident after 24 hours treatment with TNF-α (10 ng/ml) or TGF-β (1 ng/ml), as shown by phase contrast microscopy. B: The mesenchymal induction was demonstrated by expression of vimentin (IF staining, green) by the cells cultured for five days. Cortactin, predominantly expressed in epithelial cells, was stained in red; nuclei were stained by DAPI in blue. The details of morphology, as highlighted by the interior boxes, are illustrated in C. C: Lamellipodia are shown in high magnification and indicated by white arrows. D: Expression of vimentin mRNA was measured by qRT-PCT analysis of the cells treated by cytokines for 24 hours (n = 3). E: Expression of vimentin was measured by Western blot analysis. GAPDH was used as loading control. F: MMP-9 secreted into the condition media was measured by gelatinolytic zymography. G: Dermal fibroblasts constitutively express vimentin (green). TGF-β induces filamentous actin (phalloidin, red).