Fig. 4A. Intracellular localization of the p0071 domains in MCF-7 cells. (A,B) Expression of the p0071 head domain (A) and a N-terminally deleted fragment (head 149-509, B) in the pDsRed vector. Cells were Triton-extracted, PFA-fixed and stained with monoclonal desmoplakin antibody (A) or methanol-fixed and labeled with polyclonal desmoplakin antibody (B, serum 616). Plasma membrane regions denoted by arrows are shown in detail at higher magnification. (C,D) Intracellular targeting of the p0071 tail domain. Cells transfected with the p0071 tail pDsRed were fixed in methanol and stained with anti-E-cadherin (C) or desmoplakin antibodies (D). Membrane regions denoted by arrows are shown at higher magnification. (E,F,G) Intracellular localization of p0071 headless pDsRed. Cells were fixed in methanol and labeled with E-cadherin (E,F) or desmoplakin (G, serum 616) antibodies. Enlargement of membrane regions denoted by arrows shows colocalization with E-cadherin (E,F), whereas desmoplakin staining is distinct from p0071 headless localization (G). Note that desmoplakin staining is sparse along the membrane enriched in p0071 headless (G). (H,I,J) Localization of the p0071 repeats pDsRed. Cells were fixed in methanol and labeled with E-cadherin (H,I) or desmoplakin (J, serum 616) antibodies. Enlargement of membrane regions denoted by arrows shows colocalization with E-cadherin (H,I), whereas desmoplakin staining is distinct from p0071 repeat domain localization (J). Bars, 3 µm.