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. 2016 Jun 29;7:12037. doi: 10.1038/ncomms12037

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(a) FLAG-tagged Hsp90α-N-, M-, and C-domains were transiently expressed and immunoprecipitated from HEK293 cells. Co-IP of endogenous FLCN and FNIPs were assessed by immunoblotting. Empty vector (EV) was used as a control. (b) HA-tagged FNIP1 domains were transiently expressed and isolated from HEK293 cells. Co-IP of endogenous FLCN and Hsp90 were assessed by immunoblotting. Empty vector (EV) was used as a control. (c) FLAG-tagged Hsp90α domains were transiently co-expressed with HA-tagged FNIP1-D in HEK293 cells. HA–FNIP1-D was immunoprecipitated and Hsp90α–FLAG co-IP was evaluated by immunoblotting. (d) Bacterially expressed and purified FNIP1-D and Hsp90α binding affinity measured by fluorescence anisotropy. The error bars represent mean ±s.d. of three independent experiments.