Cells transfected with control vectors, PTP-MEG2, wild-type PTPN18, or different PTPN18 mutants were seeded in 96-well plates containing DMEM and 10% FBS.
For live-staining CHO cells expressing MAG or MAG-mutants were seeded onto glass cover slips.
Cells (2,000 3,000 per well) WT/K0 USP45 or rescue experiment with WT or Cys199Ala mutant were seeded into the inner of 96-well plates in eight replicates and allowed to attach for 16 h in the case of U2OS cells.
Each CHO-K1 Orexin mutant was seeded into tissue culture-treated, 384-well, black clear-bottom plates (CellBind Corning 7086), at a density of 7500 cells/well in culture medium and maintained in an incubator (5% CO2 at 37 °C) overnight.
For melan-d1 complementation assays measuring the function of myosin mutants, cells were seeded in a 24-well plate and allowed to reattach prior to infection with adenovirus.
Transiently transfected HEK-293 cells expressing C-hSERT-Y or mutants thereof were seeded on 48-well plates precoated with poly- d-lysine (0.5 × 10 cells/well) 24 h prior to the experiment.
HEK-293 transfected with WT or mutant TNFRSF1A were seeded onto slides (3 × 10) and maintained in culture for 24 hours.
The same number of pVHL null 786-O cells and 786-O cells expressing missense mutant pVHL were seeded, and their cell numbers were counted every day for 3 days (Fig. 5e).
To perform HIV-1 infection studies GHOST cells stablely expressing wild-type or mutant CXCR4 were seeded in 96-well tissue culture dishes (1.0 × 103 to 3.0 103 cells per well) and were infected with virus standardized for p24 antigen levels (NL4-3, 0,05 ng p24/well; NL-991, NL-952, 0,05 ng p24/well).
Briefly, triplicate samples of wild type B. burgdorferi or isogenic BbCRASP-2 and BbCRASP-1 mutant [25] were seeded into 1 ml tubes at a final concentration of 5×106 bacteria per ml.
