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. 2003 Aug;23(16):5790–5802. doi: 10.1128/MCB.23.16.5790-5802.2003

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Copyright © 2003, American Society for Microbiology

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FIG. 8. — HSP27, phosphorylated I-κBα, and PA700 can be found in the same cellular protein fraction. (A) Lysates from HSP27-transfected U937 cells not treated (triangles) or treated (squares) with etoposide (100 μM, 4 h) were centrifuged and fractionated with a Superose-6 column. Fractions were tested for hydrolysis of the substrate Suc-LLVY-AMC. AU, arbitrary units. (B) The presence of PA700, HSP27, and phosphorylated I-κBα (P-I-κBα) in fractions of cell extracts from nontreated (NT) and etoposide-treated (VP16) cells in the presence of MG132 (25 μM) to stabilize phosphorylated I-κBα was determined by Western blotting. (C) Immunodetection of PA700 and P-I-κBα after immunoprecipitation (IP) of HSP27 in fraction 17 of cells exposed to etoposide in the presence of MG132. ch., chain.