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. 2007 Mar 12;27(10):3750–3757. doi: 10.1128/MCB.02204-06

FIG. 2.

FIG. 2.

Mbd2 associates with the Xist gene and restores Xist repression in Mbd2-null cells. (A) Immunoprecipitation (IP) of cross-linked chromatin with an anti-Mbd2 antibody reveals that endogenous Mbd2 is associated with the CpG island (primers F3/R3) and downstream regions (primers XIST-7F/XIST-11R) of the methylated Xist gene. in, input; -ve, negative. (B) Exogenous Mbd2 is expressed from retroviral vector pBMN-ZIN Flag-Mbd2 (pBM) or empty vector pBMN-ZIN (pB) in Mbd2-null cells as detected on a Western blot using antibodies against Mbd2 or the Flag epitope tag. (C) Elevated Xist expression in Mbd2-null cells is abolished by exogenous Mbd2 expression. Levels of Xist RNA were measured by quantitative PCR analysis as described in the legend for Fig. 1B and C using primers XIST-7F/XIST-11R and XIST-4R/XIST-10F. Rel., relative. (D) Exogenous Mbd2 associates with the Xist gene in Mbd2-null cells. ChIP of cells transformed with the Mbd2-expressing vector (pBMN-ZIN Flag-Mbd2) or empty vector (pB) was done as described for panel A.