Figure 7
From: Efficient TGFβ-induced epithelial–mesenchymal transition depends on hyaluronan synthase HAS2
Knockdown of HAS2 inhibits cell migration. NMuMG cells were transfected with scrambled siRNA or siRNA against HAS2 (a) or CD44 (b), starved and then stimulated with 1 ng/ml TGFβ. Cells were then maintained in Transwell assay chambers for 24 h with TGFβ, hyaluronan or epidermal growth factor as chemotactic stimuli in the bottom chamber. Before seeding the cells in the Transwell chambers, some of the cell cultures were pretreated with CD44-blocking antibody KM114 (2.5 μg/ml) or control IgG (2.5 μg/ml) for 30 min. Transmigrating cells were stained with Giemsa and counted after bright-field images were taken. Relative number of migrating cells from a representative experiment out of three ±s.d. is shown. Asterisks indicate statistically significant differences between HAS2-depleted cells compared with those expressing HAS2 and subjected to identical treatments.
