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. 2010 Mar 10;285(19):14475–14485. doi: 10.1074/jbc.M109.084400

FIGURE 6.

FIGURE 6.

Levels of AP-1 proteins, Rb phosphorylation, and nuclear E2F in CAGE-overexpressing cells. A, protein levels of Jun and Fos subfamily members of AP-1 factors were examined in tetracycline-inducible CAGE transfectant HeLa cells by immunoblotting analysis after doxycycline (dox) treatment for 8 h. The phosphorylation level of c-Jun was evaluated by using an antibody specific to the phospho-Ser-63/Ser-73 residues of c-Jun. B, CAGE transfectant clonal cells (#2-27) were treated with or without doxycycline for the indicated time periods, and E2F-1 protein levels in the total cell lysate, the cytosolic and nuclear fractions, and the Rb immunoprecipitate (IP) were examined for by immunoblotting analysis. C, the phosphorylation level of Rb in cells (#2-27) treated with or without doxycycline was assessed by immunoblotting analysis using an antibody specific to Rb phosphorylated at the Ser-795 residue.