Figure 4

SOX2-RNAi increases TCF/LEF-signaling and decreases Nestin and OCT4 steady state protein expression levels. a: U343-MG and b: U373-MG glioma cells stably expressing TCF/LEF-driven luciferase were transduced with sh2378 or empty vector and analyzed. Note the increase in luciferase activity after knockdown of SOX2 in both glioma cell lines. Bars indicate standard deviation. * p < 0.05 when SOX2-silenced cell were compared to controls. c: Western blot analysis for stem cell markers Nestin and Oct4 in U343-MG and U373-MG cells with knockdown of SOX2 showing loss of protein expression when compared to shLuc controls. Steady state protein expression levels of astrocyte marker glial fibrillary acidic protein (GFAP) and β-Catenin remained unchanged. Equal loading of lanes was confirmed using tubulin antibodies. d: Indirect immunofluorescence showing decrease of Oct4 immunosignals in U343-MG and U373-MG cells treated with sh2378 when compared to shLuc controls. Magnification bars: 20 μm.