Fa1002

BMDMs were treated with different cytokines. Nuclear proteins were isolated with a nuclear extraction kit (Signosis, SK-0001), and analyzed by using a 96-well plate transcription factor (TF) activation array (Signosis, FA1002) according to the manufacturer’s instruction. The activity of each transcriptional factor was normalized as the fold of TFIID’s activity.

We used a 96 well plate TF activation array (# FA1002, Signosis, USA) as per the manufacturer’s guideline. MCF-7 cells were treated with NaB/ VPA for 48 hours and nuclear proteins from both control and treated cells were isolated using standard procedure and used as probe on the array plate. Transcription factor IID (TFIID) was used for normalization of readings. TFs were selected by considering the fold-change method (≤1.5 or ≥1.5) of treated over untreated cell samples. To characterize binding of various activated TFs to NIS promoter, promoter binding TF profiling array was performed (FA2002, Signosis, USA). Nuclear extracts were prepared from NaB/ VPA treated MCF-7 cells, incubated with oligo-binding mix along with NIS-promoter DNA fragment. Comparing luminescence in presence or absence of competitor human NIS promoter, binding of various TFs were predicted.