Defective dek5 chloroplast envelope membranes. (A) Relative levels of dek5 to normal chloroplast envelope proteins. Symbols indicate the type of TMD predicted for each protein. Proteins in the dotted box have only been localized to envelope membranes. (B) Time course of ³²P_i uptake in isolated normal and dek5-N961 chloroplasts. Mean and standard deviation are plotted (n = 3 biological replicates). (C) End-point ³²P_i uptake of normal and dek5-N961 chloroplast dilutions after a 5-min incubation. Mean and standard deviation are plotted (n = 3 biological replicates). Total stromal volume was estimated based on the number of chloroplasts included in each assay. (D and E) Phase-contrast micrographs of dek5-N961 (D) and normal (E) chloroplasts after a mock P_i uptake assay. Black arrows indicate intact chloroplasts. The white arrow indicates a broken chloroplast. Scale bars are 10 µm. (F) Immunoblot analysis of purified dek5-N961 and normal sibling chloroplasts. Protein was loaded on an equal OD₆₅₂ basis.