Figure 2

Wee1 siRNA treatment produces efficient target knockdown in all cell lines assayed. NCI-H1299, A549, Daoy and D54MG cells were subjected to 5 nM siRNA treatment for two days following plating. Non targeting siRNA (N1) and Wee1 siRNAs #5-#8 were all analyzed. Direct cell lysates were analyzed by Western blotting to ascertain loss of Wee1 protein levels as well as the effects on its direct target (pCDK). pCDK Western samples were spliced between H1299 and A549 to provide exposure times that result in similar levels of visible phosphorylation for all cell lines. Total CDK1 and Actin were blotted to control for total protein levels.