Tetraspanin TM4SF5 mediates loss of contact inhibition through epithelial-mesenchymal transition in human hepatocarcinoma
J. Clin. Invest. Sin-Ae Lee, et al. 118:1354 doi:10.1172/JCI33768 [Go to this article.]

Figure 4
Effects of TM4SF5 suppression on cellular morphology and cytosolic p27^Kip1 stabilization. (A) SNU449Tp cells were cotransfected with pEGFP and shTM4SF5. After 24 hours, cells were replated on coverslips for an additional 24 hours. Cells were then double-stained for p27^Kip1 and DAPI. (B) Lysates prepared in Figure 2B were immunoblotted. (C) Huh7, SNU601, and HepG2 cells with endogenous TM4SF5 were transiently transfected with scrambled or shTM4SF5, 48 hours before immunoblots. SNU449Cp and SNU449Tp cells were blotted in parallel for comparison of TM4SF5 expression. (D) Huh7 cells with endogenous TM4SF5 were transiently cotransfected with pEGFP and scrambled sequence or shTM4SF5, 48 hours before immunoblot analysis or immunostaining. Note that shTM4SF5-transfected cells have a lower cytosolic p27^Kip1 level, leading to a lower ratio of cytosolic to nucleic p27^Kip1 levels. (E) Huh7 cells stably transfected with shTM4SF5 (Huh7-shTM4SF5, Figure 2J) were transiently cotransfected with pEGFP and pcDNA3-TM4SF5. Two days later, the cells were immunostained for p27^Kip1 and DAPI. Original magnification, ×400. Data shown represent 3 isolated experiments. Scale bars: 20 μm.