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. 2006 Apr 11;395(Pt 3):449–456. doi: 10.1042/BJ20051947

Figure 5. The C-terminal KDEL motif mediates intracellular retention of Spn1.

Figure 5

(A) Immunoanalysis of supernatants and cellular extracts of COS-7 cells expressing Spn1. COS-7 cells were transiently transfected with a plasmid coding for wild-type Spn1 or for a mutant (Spn1-ΔKDEL) lacking the C-terminal KDEL motif. Supernatants and cellular extracts were collected 36 h after DNA addition, resolved by SDS/PAGE (10% gels), and analysed by Western blotting, using anti-Spn1 antibodies. (B) After immunodetection, the blotting membrane was stained with Coomassie Brilliant Blue, to check for equal loading. Lanes 1 and 2, supernatants from cells transfected with wild-type Spn1 or Spn1-ΔKDEL respectively. Lanes 3 and 4, extracts from cells transfected with wild-type Spn1 and Spn1-ΔKDEL respectively. Lanes 5 and 6, supernatants and extracts of untransfected control cells. M, molecular mass markers.