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Figure 4.
Figure 4. Functional Role of the CBP Bromodomain in CBP/p53
Association(A) Effect of CBP bromodomain on p53-induced p21
activation in 10.1 cells after UV treatment. p21 activity of the
10.1 cells transfected with p53 or p53 mutant together with p21
luciferase and β-galactosidase, with or without CBP
bromodomain, was measured in a luciferase-based assay. Mean
values of the luciferase activities represent at least three
independent cell transfections.(B) Western blotting analysis
assessing protein expression in the transfected 10.1 cells. Note
that numerals below the p21 blot represent ratio of p21
expression in the cells transfected with p53, with or without
CBP bromodomain, to that in the cells transfected with only the
empty vector pCDNA3. The signals were quantitated using Kodak 1D
Digital Image Analysis Software.(C) Assessing effects of
cotransfected bromodomains on p53-induced p21 activation in the
10.1 cells. The cell transfections and p21 luciferase activity
analysis were same as described in (A).(D) Western blots
assessing expression of various bromodomains and p53 in the
transfected 10.1 cells with or without UV-C treatment.(E) Effect
of the CBP bromodomain on cell cycle distribution induced by p53
in the 10.1 cells transfected with Us9-GFP, wild-type, or mutant
p53, with or without the CBP bromodomains. The DNA content of
the gated GFP-positive cells in G1 phase was determined by PI
staining and FACS analysis. Average values of the DNA content of
the cell cycle phases in each different experiment represent at
least three independent transfection trials.
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