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. 1999 Jan 11;144(1):21–30. doi: 10.1083/jcb.144.1.21

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ER-targeted C domain is efficiently secreted and does not interact with BiP. (A) Cells were transfected with no DNA (Mock), the ER-targeted constant region gene (ER-Cλ), or wild-type λ cDNA along with wild-type hamster BiP. Transfected cells were labeled for 2 h, culture supernatants were saved, and then lysates were prepared. The supernatants were precipitated with anti-λ (λ_m) and the lysates were divided and precipitated with either anti-rodent BiP or anti-λ. Samples were analyzed on 12% SDS-gels under reducing conditions. (B) COS-1 cells were cotransfected with one of the λ LC cDNA clones (WTλ or ER-Cλ) and either wild-type (WT) or mutant (G37) BiP. Cells were pulse-labeled as in Fig. 4 and lysed immediately in the presence of NEM and apyrase. Lysates were divided and immunoprecipitated with anti-mouse λ, anti-rodent BiP, or protein A–Sepharose alone. Samples were analyzed on 12% SDS-gels under reducing conditions.

ER-targeted C domain is efficiently secreted and does not interact with BiP. (A) Cells were transfected with no DNA (Mock), the ER-targeted constant region gene (ER-Cλ), or wild-type λ cDNA along with wild-type hamster BiP. Transfected cells were labeled for 2 h, culture supernatants were saved, and then lysates were prepared. The supernatants were precipitated with anti-λ (λ_m) and the lysates were divided and precipitated with either anti-rodent BiP or anti-λ. Samples were analyzed on 12% SDS-gels under reducing conditions. (B) COS-1 cells were cotransfected with one of the λ LC cDNA clones (WTλ or ER-Cλ) and either wild-type (WT) or mutant (G37) BiP. Cells were pulse-labeled as in Fig. 4 and lysed immediately in the presence of NEM and apyrase. Lysates were divided and immunoprecipitated with anti-mouse λ, anti-rodent BiP, or protein A–Sepharose alone. Samples were analyzed on 12% SDS-gels under reducing conditions.