After attachment, media was replaced with either fresh media (control) or media containing half-log serial dilutions of the following drugs: GDC-0941 10 nM-3000 nM-3000DC0980 (1 nM-300 nM), BKM120 (10 nM-3000 nM) and NVP-BEZ235 (1 nM-300 nM) purchased from Selleck Chemicals (Houston, TX) or in combination with bicalutamide (CDX) (SiGDC0980. Louis, MO).
LEU+ clones were grown to saturation in YPD media at 23°C to allow loss of either plasmid pTH2 or pEU42, then plated in 10-fold serial dilutions on media containing 5-fluoroorotic acid (FOA).
The selective pressure for the acquisition of compensatory mutations to restore competitive fitness is further supported by similar observations in Salmonella enterica, where the acquisition of rpoA and rpoC mutations has been shown to improve the growth rate of slow-growing rpoB mutants during serial passage in media containing RIF.
To generate dose response curves to PRO 2000 under conditions that simulate the postcoital CVL, TZM-bl cells were infected with HIV-1BaL diluted in media alone or in media containing pooled SP and mixed 1∶1 with serial dilutions of unformulated PRO 2000 diluted in pooled CVL.
The next day, growth media was replaced with treatment media containing vehicle-DMSO or paclitaxel that was serial diluted by half-log concentrations ranging from 0.3 to 30 nM.
Serial dilution of spent media containing virions is used to reinfect fresh monolayers of cells, and the extent of reinfection correlates with how many infectious virions were produced in the presence of potential inhibitor.
Yeast strains were transformed with high-copy galactose-inducible (GAL1 promoter) expression vectors containing WT human LRRK2 (residues 1300 2527) or a control empty vector, and resulting yeast clones were spotted as 5-fold serial dilutions on solid media containing glucose (LRRK2 'repressed') or galactose (LRRK2 'induced') to assess growth fitness.
Cells were plated as 10-fold serial dilutions on YPD or on media containing 5-fluoroorotic acid (FOA) then grown 3d at 23°C.
The growth properties of each mutant were tested by spotting serial dilutions of galactose-grown cells on media containing fructose or glucose as the carbon source.
Serial dilutions were spotted onto synthetic complete media containing 0.003% sodium dodecyl sulfate and 10, 25, or 50 μM MG-132 and grown at 30°.
