Tumor suppressor Tsc1 is a new Hsp90 co‐chaperone that facilitates folding of kinase and non‐kinase clients Tsc1 was immunoprecipitated from HEK293 cell.

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Presentation transcript:

Tumor suppressor Tsc1 is a new Hsp90 co‐chaperone that facilitates folding of kinase and non‐kinase clients Tsc1 was immunoprecipitated from HEK293 cell lysates using anti‐Tsc1 antibody and immunoblotted with indicated antibodies. IgG was used as control. These samples are the same as those presented in Fig A. BSA binding affinity to bacterially expressed and purified Hsp90α was determined by fluorescence anisotropy. The error bars represent mean ± SD of three independent experiments. Inorganic phosphate (Pi) standard curve. The x‐axis shows μM of Pi per assay and the y‐axis shows absorbance at 565 nm. Mean ± SD from values obtained in three independent experiments. ATPase activity of Hsp90α in vitro. Inhibitory effects of purified Tsc1‐D‐His6 on the ATPase activity of Hsp90α are shown. All the data represent mean ± SD of three biological replicates. A Student's t‐test was performed to assess statistical significance (**P < 0.005, ****P < 0.0001). Different amounts of BSA have no effect on Hsp90 ATPase activity in vitro. All the data represent mean ± SD of three biological replicates. ATPase activity of Hsp90 from five Tsc1GFAPCKO (K1–K5) mice with conditional inactivation of the TSC1 gene in glia and five Tsc1flox/+‐GFAP‐Cre and Tsc1flox/flox littermate controls (C1–C5). All the data represent mean ± SD of three biological replicates. A Student's t‐test was performed to assess statistical significance (**P < 0.005); 50 ng of the isolated Hsp90 was resolved on the SDS–PAGE gel and stained with Coomassie stain. Source data are available online for this figure. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO J, Volume: 36, Issue: 24, Pages: 3650-3665, First published: 10 November 2017, DOI: (10.15252/embj.201796700)