U343-MG and U373-MG cells with knockdown of SOX2 develop a single membrane protrusion containing ezrin and CD44. U343-MG and U343-MG glioma cells were transduced with retroviral vectors encoding EGFP and the indicated shRNA. Five days after transduction cells were fixed and stained with primary ezrin, CD44 antibodies and secondary Cy3-coupled antibodies as depicted in the images. DNA counterstaining was performed using Hoechst 33342. a, b: U343-MG cells transduced with shLuc control vector. c, d: U343-MG cells transduced with shSOX2 #sh2378. e, f: U373-MG cells transduced with shLuc control vector. g, h: U373-MG cells transduced with shSOX2 #sh2378. Note in c: and g: the development of a single cell protrusion containing ezrin and as shown in d and h the appearance of CD44 in single protrusions in cells with knockdown of SOX2. Magnification bars, in a, b, e, f: 20 μm; in c, d, g, h: 10 μm.