Immunofluorescence Staining of Salivary Gland Cells

For immunofluorescence analyses, antibodies specific for ZO-1, keratin 5, PKCζ, myosin II and phospho-myosin were purchased from BD Transduction Laboratories. Secondary antibodies included AffiniPure goat anti-mouse and goat anti-rabbit IgG Fab fragments from either Jackson ImmunoResearch Laboratories or from Molecular Probes. Localization and distribution of F-actin was followed by the staining of SMGs with Rhodamine- or Alexa-conjugated phalloidin^{3 (link),20 (link)}, while nuclei were visualized with monomeric cyanine nucleic acid stain, both from Molecular Probes. Evaluation of transfection efficiency was carried out with Cy3-siRNA, obtained from Dharmacon.