FIG. 5.
Expression of human emerin in mouse knockout cells does not detectably alter their susceptibly to HIV-1 infection. (A) Primary sequence alignment of human (h_EMD) and mouse (m_Emd) emerin proteins. *, amino acid identity; :, homology through conservative substitution as defined by reference 33. Boxed areas indicate the LEM domain, lamin A binding region, and transmembrane domain (TMD) (16). The alignment was created using ClustalW (42). (B) Western blot analysis of emerin protein expression in transformed MEF cell lines. Lane 1, protein extracted from control Emd−/y cells transduced with empty vector; lane 2, Emd−/y pLPCX-EMD cells; lane 3, Emd+/+ pLPCX cells; lane 4, HeLa-P4 cells. Human emerin migrates slightly faster than the mouse protein under these electrophoresis conditions. Migration positions of molecular mass standards are indicated to the right. (C) Normalized levels of HIV-SIN-Luc infectivities in the indicated cell types. White bars, cells infected with 2 × 106 reverse transcriptase cpm; black and gray bars, cells infected with a 10- and 100-fold dilution of virus inoculum, respectively.