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After the specified time of treatment, hot and cold wash were given followed by soaping with Lissapol D 1 g/l at 80 °C for 15 min.
The relation between surviving fraction and drug concentration is plotted to get the survival curve for breast tumor cell line after the specified time.
{text{Removal }}left( % right) = left( { 1 {text{ C}}_{text{t}} /{text{C}}_{text{o}} } right);times;100 (1) where Co is the initial concentrations of diazinon (mg/L) and Ct is the residual of diazinon (mg/L) after the specified time.
The relation between the surviving fraction and drug concentration was plotted to get the survival curve after the specified time The molar concentration required for 50%% inhibition of cell viability (IC 50 ) was calculated and compared to the reference drug 2′,7′-dichlorofluorescein (DCF).
The numbers of colonies formed were counted after the specified time interval.
After the specified time interval, cell medium was collected into 15 ml centrifuge tubes.
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For time-course experiments, the oocytes were quickly frozen in dry ice at the specified time after the last injection.
At the specified time after onset of saline or LPS infusion, animals were terminally anesthetised (i.v. sodium pentobarbital, Sagatal, Rhône Mérieux, Harlow, UK).
Supernatant and cell lysate were collected at the specified time points after chase; RSPs were immuno-precipitated using the 4E11 anti-E monoclonal antibody and then analyzed by autoradiography.
For stable cell lines, after cells migrated at 37°C for the specified time, the cells were incubated with Calcein-AM (Molecular Probes, Eugene, OR) for 1 h at 37°C before fixation.
The outcome of interest was receipt of radiation or chemotherapy within the specified time period and after the surgery date for each of the three measures.
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