FIGURE 4:

RNA interference–mediated decreased expression of ADAM12 in hepatic stellate cells induces actin cytoskeleton reorganization and loss of adhesion. HSCs were transfected with 2 nM nontargeted siRNA (Scr) or ADAM12L siRNA (siADAM12). After 48 h, Western blotting (A) and real-time PCR analyses (B) were used to confirm the efficiency and specificity of RNA interference in cell extracts. PCR controls included ADAM12S, type I collagen (Col1), and TGF-β. (C) Transfected cells were analyzed by phase-contrast microscopy (left) and stained with rhodamine-conjugated phalloidin to monitor actin stress fibers (right). Nuclei were counterstained using 4′,6-diamidino-2-phenylindole (blue). (D) Cell adhesion was evaluated by counting adherent cells vs. cells released in the medium at 24 and 48 h posttransfection. Results are expressed as the percentage of adherent cells and represented as the mean ± SD of three independent experiments (*p < 0.05; **, ^##p < 0.01). Left, Cell adhesion on plastic and collagen type I compared 24 and 48 h posttransfection. Right, cell adhesion on different substrates compared 24 h posttransfection.