Hello Amzath Nazeer a,

You serum starve the cells for 24 hours using 0.2- 0.5% FBS so that the cells survive but do not proliferate.

Serum starvation is widely used for synchronizing cells by arresting them in the G0/G1 phase of the cell cycle. After 24 hours, replace this media with serum-free media for 30 mins before you treat the cells with IFN gamma for the indicated time points.

You may categorize into 3 groups.

1. Cells treated with IFN gamma in serum free medium.

2. Cells in serum free medium without IFN gamma treatment.

3. Cells in normal growth media containing 10% FBS without IFN gamma treatment.

Please note that FBS does contain IFN gamma in the range of pg to ng.

Best.