Extracellular Sulfatases, Elements of the Wnt Signaling Pathway, Positively Regulate Growth and Tumorigenicity of Human Pancreatic Cancer Cells (original) (raw)
Figure 4
Wnt signaling in cells silenced for Sulf-2.
A: Cells were transduced with the conditional (pSico) or nonconditional lentivirus (pLVTHM) encoding either Sulf-2 specific (1413, 1143) or control (ctrl) shRNAs. The cells were lysed and equal amounts of proteins were subjected to SDS-PAGE. In case of the pSico, cells were lysed 10 days after cre transfection. Sulf-2 protein levels were determined by performing immunoblots and the same lysates were probed with anti actin antibody as a loading control. B: 100 µg total protein of the nuclear fraction from the cells were applied to SDS-PAGE and amounts of active beta-catenin were detected using an antibody against non-phosphorylated beta-catenin. As loading control, the same lysates were analyzed with an anti histone antibody. C: Parental, pLV-ctrl, pLV-1413 and pLV-1143 transduced cells were transfected with the FOP/TOPflash reporter system and luciferase activity was detected 48 hrs later. The values shown are the means±SD's for 3 independent determinations and differences between ctrl and Sulf-2 silenced cells were statistically significant (BxPC-3, p = 0.002 and (L3.6sl, p = 0.013 and 0.017 for the two silencing vectors).