American Society of Hematology

Figure 2

$Figure 2. Analyses of Mp53ERSEN and Mp53ERRES cells in vitro. (A) Immunocytochemical detection of p53ERTAM in acetone-fixed cells. Cells were counterstained with DAPI to visualize nuclei. Where indicated, cells were pretreated with 4OHT for 6 hours. Images were captured and processed as described in Figure 1C, except that the fluorescent light source was used. (B) Immunoblotting detecting p53ERTAM in whole-cell lysates and cytosolic and nuclear fractions. Actin and PCNA were used as control cytosolic and nuclear proteins, respectively. The duration of 40HT treatment was 3 hours. (C) Quantitation of the real-time RT-PCR experiment comparing expression levels of Puma and Noxa mRNAs in 40HT-treated versus untreated cells. Error bars represent SD. (D) Flow cytometric analyses of tumor cells stained with annexin V (left) and TMRE (right). Numbers refer to percentages of cells committed to apoptosis.$

Analyses of Mp53ER^SEN and Mp53ER^RES cells in vitro. (A) Immunocytochemical detection of p53ER^TAM in acetone-fixed cells. Cells were counterstained with DAPI to visualize nuclei. Where indicated, cells were pretreated with 4OHT for 6 hours. Images were captured and processed as described in Figure 1C, except that the fluorescent light source was used. (B) Immunoblotting detecting p53ER^TAM in whole-cell lysates and cytosolic and nuclear fractions. Actin and PCNA were used as control cytosolic and nuclear proteins, respectively. The duration of 40HT treatment was 3 hours. (C) Quantitation of the real-time RT-PCR experiment comparing expression levels of Puma and Noxa mRNAs in 40HT-treated versus untreated cells. Error bars represent SD. (D) Flow cytometric analyses of tumor cells stained with annexin V (left) and TMRE (right). Numbers refer to percentages of cells committed to apoptosis.

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