Hi,
I incubated the cells ,after adding the siRNA, in 5% O2. This resulted in cell death and decreased rate of proliferation in the cells. The melanoma cell line that I am working on have been cultured at 5% O2 always so to maintain the same O2 state during incubation after the knockdown, I incubated them at 5%.
Has anyone experienced the same case? I have tried the knockdown with a different melanoma cell line at the same concentration of siRNA and same duration of knockdown, at 21% O2 and it worked perfectly fine.
Hope to get suggestions on this.
Thanks for your time.
Regards,
Jyoti
The short answer is yes. 5% O2 is hypoxia for cells than ~21% at normal. Hypoxia could alter expression of your gene (and other genes) that regulate cell survival genes. Therefore, a preliminary experiment to compare expression of gene of interest at two different O2 levels could provide some evidence.
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