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. 2013 Aug 1;3(8):1315–1324. doi: 10.1534/g3.113.006692

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Copyright © 2013 Neef et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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The L189P HSF1 mutant is defective in promoting Hsp70 expression in mammalian cells. (A) hsf1^−/− MEFs were transiently transfected with an empty vector or plasmids expressing wild-type human HSF1 or the HSF1 mutants identified in this screen. The transfected cells were maintained either at 37° or heat shocked at 41° for 1 hr followed by an 8-hr recovery at 37°. Total protein extracts were analyzed for Hsp70 and HSF1 by immunoblotting. GAPDH serves as a loading control. (B) Cell lysates were prepared as in (A) and analyzed for Hsp70 levels by ELISA.