Fig. 3. Immunofluorescence localization of connexins in HeLa transfectants. HeLa (a-c), HeLa/Cx43 (d), HeLa/Cx45 (e), or HeLa/Cx46 (f) cells were cultured on glass coverslips, fixed, permeabilized, and then incubated with rabbit antiserum to either Cx43 (a,d), Cx45 (b,e), or Cx46 (c,f) PBS for 1 hour at room temperature. The cells were then washed, labeled with Cy3-conjugated anti-rabbit IgG, and photographed. HeLa cells do not express detectable Cx43 (a) or Cx46 (c), and minimal Cx45 (b). In Cx43 and Cx45 transfected cells, the transfected connexins appear largely in a linear punctate pattern at cell-cell boundaries. In contrast, Cx46 appeared as diffuse linear intercellular staining in HeLa/Cx46 cells. Scale bar: 40 µm.