Hello

I try to check the serum shock-induced change of circadian gene expression (ex Bmal1 and Per2) in BMDMs by use qPCR.

But, I have a problem, so I want to your advise.

This is my serum shock protocol.

1. BMDMs plate in 6 well plate and incubate for overnight

2. Washing with PBS and add serum free media and incubate for 2h

3. Washing with PBS and add 50% FBS-contained media and incubated for 2h

4. Washing with PBS and add 1% FBS-contained media

5. After 4, 8, 12, 16, 20 and 24h , extract the mRNA

After serum shock, I change the media to 1% FBS-contained media.

After 12h, when I saw the morphology of BMDMs , cells go to death.

I think that cell might die because nutrient deficiency.

So I want to change the media from 1% FBS-contained media to 10% FBS-contained media.

Is it okay?