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. Author manuscript; available in PMC: 2012 Aug 19.
Published in final edited form as: Cell. 2011 Aug 11;146(4):544–554. doi: 10.1016/j.cell.2011.07.006

Figure 3.

Figure 3

Pulse/Chase experiments with sequential hormone treatments. Combinations of corticosterone (cort) or estradiol demonstrate assisted loading of ER pBox protein even when assisted loading is anti-correlated with GR binding levels at later time points. Cells were pre-treated with hormone for 30 minutes, washed with hormone free media, then treated for another 15 or 30 minutes (treatment time line shown in panel C). Following fixation, (A) GFP-GR or (B) Ch-Pbox steady-state binding at the MMTV chromatin array was measured by fluorescence microscopy. Data are shown as the mean of automated measurements from 500–1000 cells per conditions; error bars denote SEM. All manipulations, including hormone addition, cell washing, cell fixation, and microscopic imaging were performed by high-throughput automated methods (see methods section for details).