Skip to main content
. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: J Cell Physiol. 2014 Mar;229(3):259–265. doi: 10.1002/jcp.24445

Figure 2.

Figure 2

Analysis of Active Chromatin Using Transcriptional Activators
  1. Images from a time series of the recruitment of the activator, Cherry-tTA-ER, to the CMV-promoter regulated transgene array in the U2OS cell line (2-6-3). Arrows indicate the location of the array. Scale bar =5 ,m. Scale bar in enlarged inset =1 ,m.
  2. (B) Schematic diagram of the synthetic transcriptional activator constructs, which can be used to analyze the effects of different transcriptional activation domains (TADs) on transcription and chromatin organization in cell lines with arrays that include the tetracycline response element repeats (Figure 1). Cherry-tTA-ER consists of the auto-fluorescent protein, Cherry, the tetracycline transcriptional activator (tTA), and the estrogen receptor hormone-binding domain (ER). tTA, itself, is a fusion between the tetracycline repressor (TetR) and the VP16 TAD. Cherry-TetR-(TAD)-ER was constructed for easy introduction of dirent TADs by directional cloning using EcoRI and KpnI. Cherry-TetR-p53 was constructed by introducing the p53 TAD into Cherry-TetR-(TAD)-ER.