Hi, I have prepared T3 (3,3′,5-Triiodo-L-thyronine sodium salt, Sigma, T6397) for cell culture according to their webpage: 1 mg of T3 is dissolved in 1 mL 1 N NaOH, diluted in 49 mL medium, filter-sterilized, aliquoted, and stored at -20c.

I have done qPCR with RNA extracted from C2C12 and P19 cells treated with 0~1 uM T3 for 24 hr. However, the expression of known responsive genes (Gpt2 and Crabp1) have not changed at all. (reference: Article Thyroid hormone regulates glutamine metabolism and anaplerot...

I would like to know if any one has any suggestion on the preparation of T3 for cell culture and how do you know if the hormone is working?

Thanks a lot