Study Design: Human umbilical vein endothelial cells were preincubated with nanomolar concentrations of estradiol, estradiol stearate, and tamoxifen.
In TFEB-eGFP localization experiments where BAPTA-AM was used, cells were preincubated with BAPTA-AM (10 µM) for 30 min and washed with fresh medium prior to drug exposure.
No increase of HL1 toxicity was detected when Jurkat, Molt-16, or CCRF-CEM cells were preincubated with C60 fullerene (data not shown).
Cells were preincubated with apoptosis inhibitors for 1 h.
For the inhibition studies, cells were preincubated with the inhibitor agonists for 30 min as indicated.
In case of CF, cells were preincubated with 2 µM of the dye [38].
Briefly, the cells were preincubated with Mitotracker Red (Invitrogen) to label the mitochondria.
Alternatively, cells were preincubated overnight with vehicle or DBT prior to incubation with dexamethasone.
In control experiments, cells were preincubated with CellTracker Green CMFDA (5-chloromethylfluorescein diacetate) at 800 nM for 10 minutes.
For this, HeLa cells were preincubated with the protein synthesis inhibitor cycloheximide for 12 h before treatment with SecTRAPs.
For eicosanoid production assessment, cells were preincubated for 24 h in serum-free DMEM followed by specific treatments.
