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. 2012 Aug 7;103(10):1788–1794. doi: 10.1111/j.1349-7006.2012.02362.x

Figure 2.

Figure 2

PDCD6 is a novel target of p53. (A) PDCD6 expression is regulated by p53. H1299 cells were transfected with Flag vector or Flag‐p53. To perform ChIP assays, choromatin‐protein complexes were immunoprecipitated with anti Flag‐p53 antibodies. Real‐time PCR amplification was performed to analyze the chromatin immunoprecitated fragments using appropriate primers for PDCD6. (B) H1299 cells were transfected with Flag vector or Flag‐p53. The mRNA level of PDCD6 was quantified by real‐time RT‐PCR. The value is normalized to GAPDH. The data represents the mean ± SD from three independent experiments, each performed in triplicate. (C) H1299 cells were transfected with Flag vector or Flag‐p53 and then analyzed by western blotting. IB, immunoblotting. (D) A549 cells and U2OS cells were treated with 1 µg/mL adriamycin (ADR) and incubated for 24 h. The mRNA level of PDCD6 was quantified by real‐time RT‐PCR. The value was normalized to GAPDH. The data represent the mean ± SD from three independent experiments, each performed in triplicate. (E) A549 cells and H1299 cells were treated with 1 µg/mL ADR for the indicated times and then analyzed by western blotting. (F) A549 cells were transfected with scramble siRNA or p53 siRNA and treated with 1 µg/mL ADR for 24 h. The mRNA level of PDCD was quantified by real‐time RT‐PCR. The value was normalized to GAPDH (left). The data represent the mean ± SD from three independent experiments, each performed in triplicate. Cell lysates were subjected to immunoblot analysis with anti‐PDCD6, anti‐p53 or anti‐tubulin (right).