Growth-arrest by serum starvation
Posted 23 March 2009 - 06:22 AM
I'm trying to investigate how does glycosylation change interaction of YY1 with its molecular partners (part of my master project).
My plan is to tranfect HEK293 cells with my constructs (calcium phosphate method) and then grow them in high-glucose. According to publication I found I should also induce growth-arrest by serum starvation when using high glucose concentration. i don't entirely understand why. If I transfect my cells and keep them in high glucose, serum free medium how would that effect proteins synthesis process? I need to investigate the role of glycosylation which is postranlational process. My plan is:
1. 0h transfect cells
2. +12h - replace medium (full medium)
3. +36h - replace medium (high glucose, serum-free)
4.48-60h conduct and experiment.
What do you think about the schedule? I wouldn't like to affect protein expression by starvation (do you know how much does starvation inhibit synthesis?). On the other hand I think 12-24 hours is enough to stimulate hehamine pathway and to glycosylate proteins.
Thanks in advance,
Posted 23 March 2009 - 02:16 PM
Posted 23 March 2009 - 02:31 PM
Thanks a lot.