Antiproliferative activity against human A549 cells assessed as inhibition of cell growth at 1.5 uM incubated for 48 hrs by MTT assay relative to control
Inhibition of ATP binding to human HSP90 alpha at 5 to 10 uM preincubated for 30 mins followed by gamma phosphate-linked ATP sepharose addition and measured after 30 mins by ATP-binding sepharose assay based western blot analysis
Inhibition of Cdc37 in human A549 cells assessed as disruption of HSP90 alpha-Cdc37 interaction by measuring decrease in Cdc37 protein level at 5 uM incubated for 6 hrs by immunoprecipitation based immunoblot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring decrease in Akt phosphorylation level at 5 uM incubated for 12 hrs by Western blot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring decrease in CDK4 protein level at 5 uM incubated for 12 hrs by Western blot analysis
Binding affinity to recombinant Cdc37 (unknown origin) assessed as inhibition of biotin labeled 3-[3-cyano-3-(4-fluorophenyl)prop-2-enoyl]oxypropyl(2R,4aS,6aR,6aS,14aS,14bR)-10-hydroxy-2,4a,6a,6a,9,14a-hexamethyl-11-oxo-1,3,4,5,6,13,14,14b-octahydropicene-2-carboxylate binding to enzyme at 0.25 to 0.5 uM incubated with compound for 2 hrs before or after incubation with biotin labeled 3-[3-cyano-3-(4-fluorophenyl)prop-2-enoyl]oxypropyl(2R,4aS,6aR,6aS,14aS,14bR)-10-hydroxy-2,4a,6a,6a,9,14a-hexamethyl-11-oxo-1,3,4,5,6,13,14,14b-octahydropicene-2-carboxylate for 2 hrs by pull down assay based immunoblot analysis
Binding affinity to full length Hsp90alpha (unknown origin) assessed as reduction in absorption at 440 nm reaching equilibrium within 5 mins at 100 uM by UV absorption analysis
Binding affinity to full length CDC37 (unknown origin) assessed as reduction in absorption at 440 nm reaching equilibrium within 15 mins at 100 uM by UV absorption analysis
Antiproliferative activity against human A549 cells assessed as inhibition of cell growth at 0.5 uM incubated for 48 hrs in presence of methyl thioglycolate by MTT assay relative to control
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring decrease in Akt phosphorylation level incubated for 12 hrs by Western blot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring decrease in CDK4 protein level incubated for 12 hrs by Western blot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring increase in BAX protein level at 1.25 to 5 uM incubated for 12 hrs by Western blot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring increase in cleaved caspase-3 protein level at 1.25 to 5 uM incubated for 12 hrs by Western blot analysis
Binding affinity to Cdc37 in human A549 cells assessed as inhibition of HSP90-Cdc37 interaction by measuring decrease in Bcl2 protein level at 1.25 to 5 uM incubated for 12 hrs by Western blot analysis
Induction of mitochondrial membrane potential loss in human A549 cells assessed as cells with depolarized mitochondria by measuring increase in J-monomer green fluorescence intensity at 0.2 uM incubated for 24 hrs by JC1 staining based fluorescence microscopic method (Rvb = 1.5%)