Journal: PLOS ONE
Article Title: Administration of novobiocin and apomorphine mitigates cholera toxin mediated cellular toxicity: Lessons from cholera toxin yeast model system
doi: 10.1371/journal.pone.0315052
Figure Lengend Snippet: A. Schematic of high-through put screening: Pre-induced cultures of BY4741/ ctxA /pGML10 and BY4741/pGML10 as a control were used. These cultures were evenly distributed in 96 well plates, then compounds from the library plate were added at the final concentrations of 20 μM. Both plates were incubated for 24 h at 30°C and were then spotted on glucose and galactose plates for readouts. B. Schematic representation of the outcome of the small molecule library screening. Structures of the few positive hits from secondary screen are shown. C. Tertiary screening of Apomorphine HCl: Pre-induced cultures of BY4741/ ctxA /pGML10 and BY4741/pGML10 as a control were spotted on solid agar plate containing galactose and galactose with apomophine (50 μM). Pictures were taken after 60–70 h D and E. Effect of Apomorphine HCl on cholera toxicity in yeast: Secondary cultures of BY4741/ ctxA -HO and vector control were set up at starting OD 600 0.05 in glucose and galactose media with different concentrations of apomorphine and were monitored for 27 hrs. OD 600 was checked every 3 h. The data shown here is collected from 3 biological triplicates (D). At the end point of liquid growth assay, the cultures were serially diluted (10 0 , 10 2 , 10 3 , 10 4 ) and were spotted on glucose and galactose plates. Pictures were taken after 60–70 hrs (E). Liquid growth assay.
Article Snippet: Spectrum collection small molecule library (MicroSource Discovery Systems Inc., Gaylordsville) was used to screen molecules against cholera toxin in yeast model. Overnight grown cultures were used to set up secondary cultures of BY4741/pGML10 and BY4741/ctxA no signal/pGML10 at A 600 nm = 0.15 in YNB-Glucose-HMU media.
Techniques: Control, Incubation, Library Screening, Plasmid Preparation, Growth Assay
