MSLN overexpression is associated with activation of NF-κB and Akt activity in PC cells. (A). Enhanced NF-κB reporter gene activity in MIA-MSLN cells. Data plotted was measured as ratio of co-transfected firefly-NF-κB reporter normalized to Actin-Renilla. ** denote p < 0.01, compared with controls, t test. (B). Nuclear protein from MIA-MSLN and control cells were subjected to western blot to detect nuclear-translocated p65, p50 and nuclear envelop protein Lamin A (panel i). Panel ii showed inhibited nuclear protein translocation when MIA-MSLN cells were transfected with MSLN specific shRNA encoding plasmid or control plasmids. (C). Whole cell extract (WCE) or nuclear protein from MIA-V and two pools of MIA-MSLN cells were probed for various IκB-α degradation pathway proteins in western blot. (D). Whole protein from MIA-V/MIA-MSLN (panel i) and from the AsPC-1 shMSLN and control cells (panel ii) were probed for various PI3k-Akt pathway proteins. Note: (panel i) data has individual lanes combined from a single electrophoresis gel.