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Appendix A. Supplementary data
Supplementary Figure 1.
Co-localization of CREB with p53 in cell nucleus. Twenty-four hours after glucose deprivation, U2OS cells were incubated with control IgG (upper panels) or with monoclonal anti-p53 and polyclonal anti-CREB URB597 (lower panels) followed by the incubation with FITC-conjugated anti-mouse IgG and rhodamine-conjugated anti-rabbit IgG. Cell nuclei were stained with DAPI. Merged image indicates the co-localization of CREB with p53 in cell nucleus.
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Supplementary Figure 2.
CREB but not CREB enhances pro-apoptotic activity of p53. (A, C) FACS analysis. U2OS cells were transfected with the constant amount of the expression plasmid for p53 together with or without the increasing amounts of CREB expression plasmid (A) or CREB expression plasmid (B). Forty-eight hours after transfection, the attached and floating cells were collected, stained with PI and the number of cells with sub-G1 DNA content was measured by FACS. (B, D) DNA ladder formation. U2OS cells were transfected as in (A) and (C). Forty-eight hours after transfection, genomic DNA was prepared and analyzed by agarose gel electrophoresis.