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. 2018 Dec 11;11:8977–8985. doi: 10.2147/OTT.S187315

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© 2018 Li et al. This work is published and licensed by Dove Medical Press Limited

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Effect of celastrol on the association of mTOR with Hsp90–Cdc37 chaperone complex.

Notes: (A) MCF7 cells were transfected with either empty vector or a vector expressing HA-tagged Hsp90α or HA-tagged Cdc37. Cell extracts were immunoprecipitated (IP) with anti-HA resin. Precipitated proteins and whole-cell extracts (WCE) were immunoblotted with anti-mTOR and anti-HA antibodies. (B) Extracts from MCF7 cells transiently transfected with either empty vector or Flag-mTOR-expression vector were immunoprecipitated with anti-Flag resin. Precipitated proteins and WCE were immunoblotted for Hsp90, Cdc37, and Flag-mTOR. (C) MCF7 cells were transfected with Flag-mTOR-expression vector for 40 hours, then treated with 5 µM celastrol for 0, 2, and 5 hours. Cell extracts were immunoprecipitated with anti-Flag resin and precipitates immunoblotted using the relevant antibodies. (D) MCF7 cells were transfected with Flag-Cdc37-expression vector for 40 hours, then treated with 5 µM celastrol for 0, 1.5, and 3 hours. Cell extracts were immunoprecipitated with anti-Flag resin and precipitates immunoblotted using the relevant antibodies.

Abbreviation: DMSO, dimethyl sulfoxide.