Phosphorylation of p73 and upregulation of PUMA in pancreatic cancer cells.

<p><b>A.</b> With or without the treatment with GO6976 or GO6976 plus NAC, lysates were immunoprecipitated with anti-p73 antibody. The immunoprecipitates were then subjected to immunoblotting using the anti-phosphorylated serine antibody. The level of the immunoprecipitates was judged by re-probing the blot with anti-p73 antibody. <b>B.</b> Total RNAs from the cells with or without treated with GO6976, GO6976 plus NAC or GO6976 plus <i>shRNA-p73</i> infection were isolated. Equal amount of RNAs was reverse-transcribed, and the expression of <i>PUMA</i> was tested by RT-PCR. <b>C.</b> With or without GO6976 treatment, MIA or PNAC-1 cells were subjected to immunoblotting analysis for PUMA expression. Equal loading of total proteins were determined by rep-probing the blot with anti-β-actin antibody.