Fig. 3.
Localization of SCIMP in tetraspanin-enriched microdomains. (A) NP-40, CHAPS, or Brij 98 lysates from Ramos cells were immunoprecipitated with anti-CD81 antibody, and samples were stained for SCIMP, MHC-II, and CD81. (B) CHAPS lysates of Ramos cells were immunoprecipitated with anti-CD37, anti-CD53, anti-CD81, or isotype control antibodies, followed by staining for SCIMP and CD81. (C and D) CHAPS (C) or Brij 98 (D) lysates of Ramos cells transfected with SCIMP-Myc or empty vector were immunoprecipitated with anti-Myc antibody, and samples were stained as indicated. (E) CHAPS lysates of human monocyte-derived dendritic cells (MDDC) immunoprecipitated with isotype control antibodies, followed by immunoprecipitation with anti-CD53 or anti-CD81, were stained for SCIMP, MHC-II, CD81, and CD53. (F) Spontaneously polarized Ramos cells transfected with SCIMP-GFP were stained for CD37, CD53, CD81, or control CD45 and analyzed by confocal microscopy. Scale bar = 5 μm. Data are representative of two (A and F) or three (B, C, and D) independent experiments.