Accumulation and ubiquitination of insoluble proteins in the <i>chip</i>, <i>nbr1</i> single and double mutants under heat stress.

<p>(A) Accumulation of insoluble proteins. Leaf tissues from wild-type (WT), <i>chip, nbr1</i> single and double mutants collected at indicated hours (h) under 45°C for preparation of total, soluble and insoluble proteins as described in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004116#s4" target="_blank">Materials and Methods</a>. Total proteins in the starting homogenates and insoluble proteins in the last pellets were determined for the percentages of insoluble proteins to total proteins were calculated. (B) Ubiquitination of insoluble protein aggregates in wild type (WT), <i>chip, nbr1</i> single and double mutants collected at indicated hours (h) under heat stress. Proteins from the last pellets were subjected to SDS-PAGES and probed with anti-ubiquitin monoclonal antibody. The experiment was repeated three times with similar results.