For washout, cells were washed in PBS three times and resuspended in complete RPMI.
The MSP results were challenged by cytological analysis during the post-surgical monitoring: 5/12 patients showed positive cytology at the time of cell collection to MSP analysis, but only one of these exhibited RARB hypermethylation in the same washout cells.
RASSF1A gene MSP analysis in washout cells showed discordant results since its hypermethylation was not detected in 4 recurrent cases, although 2 negative cases for tumor cells using cytology showed this tumor tag, which suggest that these patients are under high risk for tumor recurrence.
After Aplidine-washout, cells which were in the S phase (BrdUrd positive cells) during drug treatment, progressed through this phase of the cell cycle more slowly than control cells.
Results indicated that the post-washout proliferation rates of both eribulin- and 5-FU-pretreated cells were slightly slower at 2 days, but had recovered to virtually identical rates as DMSO control cells by day 4, indicating that all post-washout cells injected into tail veins retained full viability and proliferative capacity (Supplementary Figure S2).
One hour after brefeldin A washout, increased cell surface expression of both antigens with marked cell surface co-localization was observed.
In some experiments, MEF were seeded and treated with 332 nM nocodazole for 18 hours and following washout, cells were incubated in growth medium or growth medium with mAb B1C3 and P1C9 (10 µg/ml) or mIgG (10 µg/ml) for 3 or 6 hours.
21 hr after history washout, cells were diluted into the perturbation environment and grown for 12 hr.
92 hr after history washout, cells at 2, 4, and 8 ng/ml ATc were exposed to an osmotic stress (600 mM NaCl).
92 hr after history washout, cells at different distances from the phenotypic switch were exposed to an increase in (IPTG) from 40 μM to 360 μM.
