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. 2008 Oct 6;183(1):87–99. doi: 10.1083/jcb.200803034

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© 2008 Wang et al.

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.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 1. — Arabidopsis ppi2 homozygous plants expressing atToc159 and atToc159-A864R. Heterozygous ppi2 plants were transformed with CaMV35S∷atToc159 and CaMV35S∷atToc159-A864R binary vectors encoding atToc159 and atToc159-A864R, respectively. (A) Homozygous ppi2 plants carrying the CaMV35S∷atToc159-A864R transgene (A864R:ppi2) were identified by PCR genotyping using primers specific for the ppi2 allele or the wild-type atTOC159 gene (atToc159 WT). (B) Serial dilutions of protein extracts from A864R:ppi2, WT:ppi2, and wild-type (WT) plants were immunoblotted with anti-atToc159 serum. (C) Isolated chloroplasts (T) from wild-type and A864R:ppi2 plants were extracted with alkaline carbonate, pH 11.5, and separated into membrane pellet (P) and soluble (S) fractions. The fractions were resolved by SDS-PAGE and immunoblotted with anti-atToc159 serum. (D) Detergent-solubilized chloroplasts from WT:ppi2 and A864R:ppi2 plants were subjected to immunoprecipitation with anti-atToc159 serum. 10% of the total extracts (S) and unbound fractions (F) or the total fractions from the immunoprecipitates (IP) were resolved by SDS-PAGE and immunoblotted with antisera to atToc159, atToc75, and atToc33.