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. 2003 Apr;23(8):2645–2657. doi: 10.1128/MCB.23.8.2645-2657.2003

FIG. 7.

FIG. 7.

Putative activation mechanisms by MBD2a. (A) HEK-293 cells were transiently transfected with HA-RHA, HA-MBD2a, and HA-MBD2b. The cell lysates were immunoprecipitated with anti-HA antibody and immunoblotted with anti-Pol II antibody (upper panel) or anti-HA antibody (lower panel). The asterisk indicates immunoprecipitated IgG heavy chain. (B) Ch-IP assay of D5 cells containing integrated copies of the rat somatostatin gene. D5 cells were transfected with HA-RHA, HA-MBD2a, and HA-MBD2b and cotransfected with PKAmut or PKAwt. After 24 h of transfection, formaldehyde cross-linked protein-DNA complexes were immunoprecipitated with anti-HA. A PCR analysis to amplify somatostatin promoter sequences (−100 to +55) is shown, including input DNA. (C) RT-PCR of total RNA from D5 cells expressing chromosomal copies of the rat somatostatin gene (som). Cells were transfected with HA-RHA, HA-MBD2a, and HA-MBD2b and cotransfected with PKAmut or PKAwt. The expression of β-actin, a ubiquitous mRNA, was used as an internal control.