Figure 4
From: Efficient TGFβ-induced epithelial–mesenchymal transition depends on hyaluronan synthase HAS2
Cellular localization of HAS2 and hyaluronan. NMuMG cells were transfected with scrambled siRNA or siRNA against HAS2 (b and c), starved, and stimulated with 1 ng/ml TGFβ for 12 h (a) or 48 h (a–c). Cells were fixed and immunostained with anti-HAS2 (green in a). Hyaluronan was visualized by biotinylated G1 probe (green in b and c), Actin filaments were visualized by TRITC-phalloidin (red) and cell nuclei with DAPI (blue). To visualize extracellular hyaluronan (b), cells were not permeabilized and to visualize intracellular hyaluronan (c), cells were first treated with 10 U/ml Streptomyces hyaluronidase to remove extracellular hyaluronan, washed extensively and then permeabilized. Arrows indicate plasma membrane protrusions stained for HAS2 or hyaluronan. A representative experiment out of two is shown.
