Fig. 5
TC2N blocks Cdk5-mediated p53 phosphorylation by promoting the degradation of Cdk5. a H460 cells were transfected with negative control or TC2N shRNA for 48 h and then cells were lysed and were subjected to WB using anti-phospho-p53 antibodies. b H460 cells were co-transfected with negative control or TC2N shRNA and Cdk5 siRNA. At 48 h after transfection, the cells were lysed and were subjected to WB using indicated antibodies. c Lysates from A549 vector control or TC2N cells treated with 20 µM cycloheximide (CHX) for the indicated times were subjected to WB (Up). Relative Cdk5 levels were quantified by ImageJ software (Down). d Lysates from A549 vector control or TC2N cells were treated with 20 µM MG132 for the indicated times were subjected to WB. e A549 vector control or TC2N cells were transfected with 7.5 µg of ubiquitin-expressing plasmids. At 24 h after the transfection, cells were treated 20 µM MG132 for 24 h. Cell lysates were IP with anti-Cdk5 antibody and analyzed by WB with anti-ubiquitin antibody, and normal IgG was used as a negative control. Whole-cell lysates were used as a positive control (Input)
