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. 2009 Apr 21;284(24):16135–16145. doi: 10.1074/jbc.M109.007484

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 8. — OTUB1 stabilizes ERα in the chromatin. A, OTUB1 is located in the insoluble chromatin fraction in Ishikawa cells. The nuclear fraction was isolated from untreated Ishikawa cells and subfractioned to insoluble chromatin fraction and a soluble nuclear fraction as described under “Experimental Procedures.” The fractions were analyzed by Western blot with indicated antibodies. B, expression of OTUB1 in Ishikawa cells causes retention of ERα in the chromatin. Ishikawa cells were transfected with pCMV6 or HA-OTUB1 and treated as indicated with MG132 (4 μm) for 4 h and with estradiol (E2) or ethanol (EtOH) for 2 h. The chromatin fractions were isolated as in A and analyzed using indicated antibodies. The protein bands were quantified using Image J and normalized to histone. IB, immunoblot.