Figure 5. Reorganization of SBSN chromatin structure upon BORIS induction.

BORIS expression was induced by 0, 0.0313 or 1 µg/ml doxycycline 24 hours after transfection with control empty vector or with BORIS expressing vector. Cell nuclei were isolated and digested with MNase. Purified DNA after 8 min of digestion with MNase (Figure S6) was used in PCR for primer extension experiments as described in the Methods. Relative localization of the TSS and region of chromatin reorganization is indicated. DNA digestion outside of SBSN promoter was used as loading control. Note: DNA was most accessible for MNase digestion at 0 µg/ml doxycycline, even at overall underloading of this sample. M – Hi-Lo DNA ladder (BioRad).