Fig 1.
In-vivo phenotypic study demonstrating effects of 20-hydroxyecdysone on adult worms.
(A) Graph showing recovered parasites from the study animals after necropsy. (B) Control worm showing normal length of the adult female parasite. (C) Ecdysone treated worm showing stunted growth and shorter length.
Fig 2.
Performance of the mammalian cell assay.
Results of a typical assay carried out in a 96 well format are shown. Columns indicate the mean and error bars the standard deviation of six control (uninduced) and six experimental (induced) wells. (A) Assay conducted with the transiently transfected cells with 150ng per well of each construct (BmaEcR- GAL4, RXR-VP16, GLuc reporter) using Lipofectamine, and treated with 10μM 20-hydroxyecdysone for 48 hrs. (B) Assay conducted with the stably transfected cells.
Fig 3.
Compounds identified in the screen of compounds active against the BmaEcR.
Cells were treated with 10μM of each drug for 48 hrs. The blue bars represent all other compounds tested; 20E is represented as green bar and the red bar indicates the activity present in untreated cells. All columns represent the mean and the error bars the standard deviation in triplicate wells. The horizontal line indicates the activity expected for compounds with no activity.
Fig 4.
EC50 values and structures of compounds identified in the preliminary screen of compounds interacting with the BmaEcR using the cell-based luciferase assay.
Fig 5.
Cartoon depiction of the B. malayi EcR LBD homology model.
The secondary structural element are colored in purple. The red dotted circle denotes the active site for the docking studies.
Fig 6.
Predicted structures of agonists docking in the active site of the homology model of the BmaEcR LBD.
(A-D) The panels show 20-hydroxyecdysone, ponasterone A, muristerone A and diacylhydrazine #28 binding into the active site.
Table 1.
XP descriptor analysis given from XP docking of hormones and agonists in B. malayi EcR-LBD binding site.
Fig 7.
Structure and EC50 values of the compounds predicted to be BmaEcR ligands using the in-silico virtual screening method.
Fig 8.
In-vitro phenotypic study demonstrating effect of Ponasterone A, Muristerone A and 20E on adult female worms.
The bars depict counts of progeny expelled per ml of media. 1 and 2 represent two biological replicates of the treated and control for the expulsed progeny. The error bars denote the standard deviation of the technical replicates. The colored bars represent each consecutive day of the experiment.