Fig. 5. Recruitment of RII onto Reh chromatin in mitotic extract derived from Reh-RII, but not Reh-RII(T54E), cells. Purified Reh nuclei were incubated for 2 hours in extract derived from mitotic Reh, Reh-RII(T54E) or Reh-RII cells to elicit chromatin condensation. (A) Localization of AKAP95 (red) and RII (green) was assessed by immunofluorescence analysis of condensed chromatin. Bar, 10 µm. (B) Condensed chromatin masses were recovered by sedimentation through sucrose and proteins immunoblotted using anti-AKAP95 and anti-RII mAbs. (C) AKAP95 and RII were immunoprecipitated from each condensed chromatin fraction after solubilization with MNase, and immune precipitates were immunoblotted using anti-pT antibodies.