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Fig. 5. Possible mechanisms for the reduction in Ring1B binding upon Oct3/4
depletion or differentiation cues. (A) Western blot demonstrating
changes in the levels of Oct3/4, Ring1B, Phc1, Eed, Suz12 and H3K27me3 after
conditional deletion of Oct3/4 by tetracycline (Tc) treatment of
ZHBTc4 ES cells. (B) Changes in gene expression levels for Eed,
Suz12, Ezh2, Ring1B, Phc1 and Bmi1 after tetracycline treatment
(+Tc) of ZHBTc4 ES cells were determined by real-time PCR, normalized to an
Actb control and depicted as fold changes relative to the
tetracycline-untreated (-Tc) ES cells. Error bars are based on the s.d.
derived from triplicate PCR reactions. (C) Physical interaction of
Ring1B and Oct3/4 in wild-type ES cells demonstrated by reciprocal
immunoprecipitation/immunoblot analyses. Antibodies used for
immunoprecipitation (IP, top) and immunoblotting (IB, side) are indicated. The
association between Ring1B and Oct3/4 proteins remains intact in the presence
of ethidium bromide (+EtBr), a DNA-intercalating drug that can disassociate
proteins from DNA.
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