And so goes the rationalization for anyone who has ever shopped at Wal-Mart, mocking the type of people who shop at Wal-Mart.
ZE cells were transfected with 1, 2, or 3 μg of mock, wild-type, S270A, or S270E mutant nSMase1 DNA.
Viability decreased in cells transfected with the S270E mutant but not in those transfected with mock, wild-type, or S270A mutant.
The cells were transfected with the nSMase1 mock, wild-type, S270A mutant, and S270E mutant constructs using TransIT Jurkat Transfection Reagent (Mirus Bio LLC, Madison, WI, USA).
Viability decreased in cells transfected with the S270E mutant but not in those transfected with the mock, wild-type, or S270A mutant (Supplementary Figure S8e).
This FRET biosensor was stably co-transfected either with empty vector (mock), wild-type Taspase1 (WT) or dnTASP1 (DN) into HEK293 cells.
Viability decreased in cells transfected with the S270E mutant but not in those transfected with mock, wild-type, or the S270A mutant.
When we transiently overexpressed the mock, wild-type, S270A, and S270E mutants of nSMase1 in ZE cells, cells overexpressing S270E exhibited higher nSMase activity and ceramide levels than those overexpressing the mock mutants.
After treatment with z-VAD-fmk for 1 h prior to transfection, the mock, wild-type, H272A, S270A, and S270E mutant constructs were transiently transfected into Jurkat T cells.
Briefly, various amounts of biotinylated TGF-β1 (2.5 60 ng) were added to 10 HEK-293T cells that were transiently transfected with mock, wild-type or N70/94Q versions of TβRII.
