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

Figure 7. Overexpression of FNIPs enhance Hsp90 binding to drugs.

Figure 7

(a) Structure of human Hsp90α N-domain bound to ATP (PDB: 3T0Z). (b) HA–FNIP1, HA–FNIP2 or empty vector (EV) were transiently overexpressed in HEK293 cells. Lysates were incubated with ATP agarose. Hsp90 binding to ATP agarose was examined by immunoblotting. (c) Structure of human Hsp90α N-domain bound to GB (PDB: 3TUH). (d) cMyc–FNIP1, cMyc–FNIP2 or empty vector (EV) were transiently overexpressed in HEK293 cells. Lysates were incubated with indicated amounts of biotinylated GB followed by streptavidin agarose beads. Hsp90 was detected by immunoblotting. (e) Structure of human Hsp90α bound to related SNX2112 compound, tetrahydro-4H-carbazol-4-one (PDB: 3D0B). (f) Lysates from d were incubated with indicated amounts of biotinylated SNX2112 followed by streptavidin agarose beads. Hsp90 was detected by immunoblotting. (g) FNIP1 and FNIP2 were silenced by siRNA in HEK293 cells. Lysates were incubated with ATP agarose. Hsp90 binding to ATP agarose was examined by immunoblotting. NT represents non-targeting siRNA control pools. (h) Lysates from g were incubated with indicated amounts of biotinylated GB followed by streptavidin agarose beads. Hsp90 was detected by immunoblotting.