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Figure 3 | Virology Journal

Figure 3

From: Repressor element-1 silencing transcription factor/neuronal restrictive silencer factor (REST/NRSF) can regulate HSV-1 immediate-early transcription via histone modification

Figure 3

Episomal plasmids remained in extra-chromosomal form and associated with nucleosomes in stable cells. A. Analysis of nucleosomal formation on transient transfected reporter plasmid. Plasmid pICP4 was cotransfected with control vector (Lane 1, 2, 7, and 8), pFLAG-REST (Lane 3, 4, 9, and 10), and pCMVp73 (Lane 5, 6, 11, and 12). Lane 1–6: Ethidium bromide staining. Lane 7–12: plasmid-specific nucleosmal protected ladder detected by vector probe. Lane 13 and 14: Naked plasmid digestion control. B. Analysis of nucleosomal formation on episomal plasmids. The nuclei from stable 293HEK-pICP22 were subjected to MNase digestion followed by Southern blot hybridization. Lane 1–3: ethidium bromide staining of the genomic DNA nucleosomal protected ladder. Lane 4–6: episomal plasmid-specific nucleosmal protected ladder detected by vector probe. C. Examination of episomal status of plasmids in the stable cells harboring pICP4 by Southern hybridization. Lane 1. Total DNA purified from 1.28 × 106 stable cells; Lane 2: 10 pg plasmid; Lane 3: 0.1 ng plasmid DNA; 4: 1 ng plasmid DNA; 5: 10 ng plasmid DNA.

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Virology Journal

ISSN: 1743-422X