Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 1998 May 18;141(4):1061–1071. doi: 10.1083/jcb.141.4.1061

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

(A) In vitro translation of mouse Eva transcript. In vitro translation was performed in the absence (lanes 1, 3, and 5) or in the presence (lanes 2, 4, and 6) of canine pancreatic microsomal membranes. Translated polypeptides were fractionated through a 12% SDS-PAGE gel and then analyzed by fluorography. Lanes 1 and 2: no RNA as negative control. Lanes 3 and 4: Saccharomyces cerevisiae α-factor mRNA, used as positive control for both translation and glycosylation. The α-factor and the core-glycosylated precursor have molecular weights of 18.6 and 32 kD, respectively. Lanes 5 and 6, Eva mRNA. Lanes 7 and 8: Eva mRNA as in lane 6 or treated with the enzyme endo-H. Positions of molecular mass markers (in kDs) are indicated on the right. (B) Cellular localization and immunoblot of chimeric EVA. Left, anti-myc immunofluorescence on CHO cells transfected with either pcDNA.3 vector (top) or Eva.myc vector (bottom). Right, anti-myc immunoblot on Triton X-100 lysates from the same cells. (C) Anti-EVA immunoblot. Western analysis with rabbit anti-EVA serum of Triton X-100 lysates from CHO cells transfected with either pcDNA.3 vector or Eva.myc vector and lysates in SDS of thymic epithelial cell lines A89A and TNC.R3.1. (D) EVA recovery in insoluble cell fraction. TNC.R3.1 and A89A cells were lysed in hypotonic buffer and separated into three fractions: P1, pellet of low-speed centrifugation; P100, pellet recovered after 100,000 g centrifugation, S100, soluble fraction. Equivalent amounts of all samples were resolved by SDS-PAGE in a 5–15% gradient gel and immunoblotted with rabbit anti-EVA antiserum, anti-actin and anti-src mAbs. EVA is exclusively present in the P1 fraction of TNC.R3.1 cells.