Ecdysone Response Timecourse in Kc Cells

Kc167 Cells were plated at 0.5x106/ml and grown at 23.50C. They were treated with 20-HE (Sigma) at a final concentration of 5x10-7M. The reference sample was total RNA isolated at 0 hr. after treatment. The experimental samples were total RNA isolated from cells treated for 1, 3, 6, 12, 24, and 48 hours. Samples were taken at 0hr, 1hr, 3hr, 6hr, 12hr, 24hr and 48hr. Briefly, cells were spun down and Total mRNA was extracted with OligodT (Ambion) nd reverse transcribed in the presence of oligodeothymidine and random hexamers and labeled to the whole genome in-house cDNA microarray and ecdysone responsive genes were identified. Ecdysone responsive genes were identified at each timepoint using a Limma t-test.